PMID- 15042592 TI - Isolation of an acetyl-CoA synthetase gene (ZbACS2) from Zygosaccharomyces bailii. AB - A gene homologous to Saccharomyces cerevisiae ACS genes, coding for acetyl-CoA synthetase, has been cloned from the yeast Zygosaccharomyces bailii ISA 1307, by using reverse genetic approaches. A probe obtained by PCR amplification from Z. bailii DNA, using primers derived from two conserved regions of yeast ACS proteins, RIGAIHSVVF (ScAcs1p; 210-219) and RVDDVVNVSG (ScAcs1p; 574-583), was used for screening a Z. bailii genomic library. Nine clones with partially overlapping inserts were isolated. The sequenced DNA fragment contains a complete ORF of 2027 bp (ZbACS2) and the deduced polypeptide shares significant homologies with the products of ACS2 genes from S. cerevisiae and Kluyveromyces lactis (81% and 82% identity and 84% and 89% similarity, respectively). Phylogenetic analysis shows that the sequence of Zbacs2 is more closely related to the sequences from Acs2 than to those from Acs1 proteins. Moreover, this analysis revealed that the gene duplication producing Acs1 and Acs2 proteins has occurred in the common ancestor of S. cerevisiae, K. lactis, Candida albicans, C. glabrata and Debaryomyces hansenii lineages. Additionally, the cloned gene allowed growth of S. cerevisiae Scacs2 null mutant, in medium containing glucose as the only carbon and energy source, indicating that it encodes a functional acetyl-CoA synthetase. Also, S. cerevisiae cells expressing ZbACS2 have a shorter lag time, in medium containing glucose (2%, w/v) plus acetic acid (0.1-0.35%, v/v). No differences in cell response to acetic acid stress were detected both by specific growth and death rates. The mode of regulation of ZbACS2 appears to be different from ScACS2 and KlACS2, being subject to repression by a glucose pulse in acetic acid-grown cells. PMID- 15042593 TI - GFP as a quantitative reporter of gene regulation in Candida albicans. AB - A system has been developed for the quantitative analysis of gene expression within individual Candida albicans cells in infected tissue. The system is based on the plasmid pGFP, which contains the codon-optimized yeast enhanced green fluorescent protein (yEGFP; Cormack et al., 1997) cloned between a basal CaADH1 promoter and the ScCYC1 terminator on an integrating vector. Promoters were inserted into pGFP and GFP levels measured in individual cells by quantitative fluorescence microscopy. Analysis of pPCK1-GFP and pMET3-GFP fusions revealed that GFP folds rapidly following gene induction, and is turned over rapidly following gene repression. Hence, single cell fluorescence measurements are likely to reflect ongoing gene expression levels with reasonable accuracy. pACT1 GFP expression levels were relatively constant during growth of C. albicans in both yeast and hyphal forms, and during growth in vivo in the mouse model of systemic infection. Therefore, pACT1-GFP provides a useful control for this quantitative GFP-based system in future analyses of C. albicans molecular responses during fungal infections. PMID- 15042594 TI - Alcohol dehydrogenases that catalyse methyl formate synthesis participate in formaldehyde detoxification in the methylotrophic yeast Candida boidinii. AB - Methyl formate synthesis during growth on methanol by methylotrophic yeasts has been considered to play a role in formaldehyde detoxification. An enzyme that catalyses methyl formate synthesis was purified from methylotrophic yeasts, and was suggested to belong to a family of alcohol dehydrogenases (ADHs). In this study we report the gene cloning and gene disruption analysis of three ADH encoding genes in the methylotrophic yeast Candida boidinii (CbADH1, CbADH2 and CbADH3) in order to clarify the physiological role of methyl formate synthesis. From the primary structures of these three genes, CbAdh1 was shown to be cytosolic and CbAdh2 and CbAdh3 were mitochondrial enzymes. Gene products of CbADH1, CbADH2 and CbADH3 expressed in Escherichia coli showed both ADH- and methyl formate-synthesizing activities. The results of gene-disruption analyses suggested that methyl formate synthesis was mainly catalysed by a cytosolic ADH (CbAdh1), and this enzyme contributed to formaldehyde detoxification through glutathione-independent formaldehyde oxidation during growth on methanol by methylotrophic yeasts. PMID- 15042595 TI - A new approach to species determination for yeast strains: DNA microarray-based comparative genomic hybridization using a yeast DNA microarray with 6000 genes. AB - DNA-DNA hybridization is known as the superior method in the elucidation of relationships between closely related taxa, such as species and strain. For species determination we propose a new DNA-DNA hybridization method: the DNA microarray-based comparative genomic hybridization (CGH) method, using a yeast DNA microarray with approximately 6000 genes. The genome from a yeast strain as a sample strain (Sample) was labelled with Cy3-dye and hybridized to a single DNA microarray, together with the Cy5-labelled genome of S. cerevisiae S288C as a reference strain (Reference). The log2 ratio values [log2[Cy3(Sample)/Cy5(Reference)]: Ratio] of signal intensities of all the gene spots were estimated and divided into the following groups: Ratio < or = -1; -1 < Ratio < 1; 1 < or = Ratio. The hybridization profiles of the genomes of type strains belonging to the genus Saccharomyces were significantly different from that of S. cerevisiae S288C. The Ratio-based grouping allowed us to discriminate between some species from S. cerevisiae more clearly. Furthermore, cluster analysis discriminated between closely related species and strains. Using this method, we were able to not only perform species determination but also to obtain information on alternation in gene copy number of such gene amplifications and deletions with single-gene resolution. These observations indicated that DNA microarray-based CGH is a powerful system for species determination and comparative genome analysis. PMID- 15042596 TI - The Saccharomyces cerevisiae alcohol acetyl transferase Atf1p is localized in lipid particles. AB - The yeast alcohol acetyl transferase I, Atf1p, is responsible for the major part of volatile acetate ester production in fermenting Saccharomyces cerevisiae cells. Some of these esters, such as ethyl acetate and isoamyl acetate, are important for the fruity flavours of wine, beer and other fermented beverages. In order to reveal the subcellular localization of Atf1p and further unravel the possible physiological role of this protein, ATF1::GFP fusion constructs were overexpressed in brewer's yeast. The transformant strain showed a significant increase in acetate ester formation, similar to that of an ATF1 overexpression strain, indicating that the Atf1p-GFP fusion protein was active. UV fluorescence microscopy revealed that the fusion protein was localized in small, sphere-like organelles. These organelles could be selectively stained by the fluorescent dye Nile red, indicating that they contained high amounts of neutral lipids and/or sterols, a specific characteristic of yeast lipid particles. Purification of lipid particles from wild type and ATF1 deletion cells confirmed that the Atf1p GFP fusion protein was located in these organelles. Furthermore, a clear alcohol acetyl transferase activity could be measured in the purified lipid particles of both wild type and transformed cells. The localization of Atf1p in lipid particles may indicate that Atf1p has a specific role in the lipid and/or sterol metabolism that takes place in these particles. PMID- 15042597 TI - Androgen receptor CAG repeat length correlates with semen PSA levels in adolescence. AB - BACKGROUND: Androgens exert their action through the androgen receptor (AR). The length of the AR CAG repeat is inversely correlated to receptor function and short CAG length might be a risk factor for development of prostate cancer. Our aim was to investigate whether CAG repeat number might have an impact on prostate function in adolescence. METHODS: AR genotyping was performed by direct sequencing of leukocyte DNA from 274 military conscripts. All men underwent endocrine evaluation and semen analysis. RESULTS: PSA in seminal plasma, total sperm count and motility all are inversely correlated with CAG numbers (rho = 0.128, P = 0.038; rho = -0.156, P = 0.010; rho = -0.158, P = 0.011), whereas serum levels of free testosterone (rho = 0.132; P = 0.029) and luteinizing hormone (rho = 0.126; P = 0.037) are positively correlated to CAG length. No correlation between seminal PSA and serum testosterone, neither free nor total, was found. CONCLUSIONS: In adolescence, AR genotype, but not serum testosterone, is associated with the level of seminal PSA. PMID- 15042598 TI - Loss of BMP2, Smad8, and Smad4 expression in prostate cancer progression. AB - BACKGROUND: The role of the bone morphogenetic protein (BMP) pathway in prostate cancer (PC) is unclear. This study aimed to characterize aspects of the BMP pathway in PC by assessing BMP2, Smad8, and Smad4 expression in normal, hyperplastic, and malignant prostate tissue, and to correlate findings with progression to PC. METHODS: Radical prostatectomy (RP) specimens from 74 patients with clinically localized PC (median follow-up 51 months, range 15-152), 44 benign prostatic hypertrophy (BPH) lesions, and 4 normal prostates (NPs) were assessed for BMP2, Smad8, and Smad4 expression using immunohistochemistry. RESULTS: Both BMP2 (P < 0.001) and nuclear Smad4 (P < 0.0001) expression were significantly decreased in PC compared to benign prostate tissue. Nuclear Smad8 was present in normal/benign prostate tissue but absent in PC and adjacent hyperplasia. Furthermore, loss of BMP2 (P < 0.001) and decreased nuclear Smad4 (P = 0.05) expression correlated with increasing Gleason score. CONCLUSIONS: These data suggest that decreased BMP2, nuclear smad8 and nuclear Smad4 expression are associated with the progression to PC, and in particular loss of BMP2 and Smad4 are related to progression to a more aggressive phenotype. PMID- 15042599 TI - Analysis of vitamin D-regulated gene expression in LNCaP human prostate cancer cells using cDNA microarrays. AB - BACKGROUND: 1,25-dihydroxyvitamin D(3) [1,25(OH)2D3] exerts growth inhibitory, pro-differentiating, and pro-apoptotic effects on prostate cells. To better understand the molecular mechanisms underlying these actions, we employed cDNA microarrays to study 1,25(OH)2D3-regulated gene expression in the LNCaP human prostate cancer cells. METHODS: mRNA isolated from LNCaP cells treated with vehicle or 50 nM 1,25(OH)2D3 for various lengths of time were hybridized to microarrays carrying approximately 23,000 genes. Some of the putative target genes revealed by the microarray analysis were verified by real-time PCR assays. RESULTS: 1,25(OH)2D3 most substantially increased the expression of the insulin like growth factor binding protein-3 (IGFBP-3) gene. Our analysis also revealed several novel 1,25(OH)2D3-responsive genes. Interestingly, some of the key genes regulated by 1,25(OH)2D3 are also androgen-responsive genes. 1,25(OH)2D3 also down-regulated genes that mediate androgen catabolism. CONCLUSIONS: The putative 1,25(OH)2D3 target genes appear to be involved in a variety of cellular functions including growth regulation, differentiation, membrane transport, cell-cell and cell-matrix interactions, DNA repair, and inhibition of metastasis. The up regulation of IGFBP-3 gene has been shown to be crucial in 1,25(OH)2D3-mediated inhibition of LNCaP cell growth. 1,25(OH)2D3 regulation of androgen-responsive genes as well as genes involved in androgen catabolism suggests that there are interactions between 1,25(OH)2D3 and androgen signaling pathways in LNCaP cells. Further studies on the role of these genes and others in mediating the anti cancer effects of 1,25(OH)2D3 may lead to better approaches to the prevention and treatment of prostate cancer. PMID- 15042600 TI - Purification of the keratan sulfate proteoglycan expressed in prostatic secretory cells and its identification as lumican. AB - BACKGROUND: Secretory epithelial cells of human prostate contain a keratan sulfate proteoglycan (KSPG) associated with the prostatic secretory granules (PSGs). The proteoglycan has not been identified, but like the PSGs, it is lost in the early stages of malignant transformation. METHODS: Anion exchange and affinity chromatography were used to purify KSPG from human prostate tissue. Enzymatic deglycosylation was used to remove keratan sulfate (KS). The core protein was isolated using 2D gel electrophoresis, digested in-gel with trypsin, and identified by peptide mass fingerprinting (PMF). RESULTS: The purified proteoglycan was detected as a broad smear on Western blots with an apparent molecular weight of 65-95 kDa. The KS moiety was susceptible to digestion with keratanase II and peptide N-glycosidase F defining it as highly sulfated and N linked to the core protein. The core protein was identified, following deglycosylation and PMF, as lumican and subsequently confirmed by Western blotting using an anti-lumican antibody. CONCLUSIONS: The KSPG associated with PSGs in normal prostate epithelium is lumican. While the role of lumican in extracellular matrix is well established, its function in the prostate secretory process is not known. It's potential to facilitate packaging of polyamines in PSGs, to act as a tumor suppressor and to mark the early stages of malignant transformation warrant further investigation. PMID- 15042601 TI - Modulation of anti-adhesion molecule MUC-1 is associated with arctiin-induced growth inhibition in PC-3 cells. AB - BACKGROUND: Lignans have been reported to possess anti-tumor activity in various cancer cells. However, their anticancer effects in human prostate cancer have not been well established. Here, we examine the effect of arctiin, a lignan compound, on growth regulation in prostate cancer PC-3 cells. We postulated that arctiin modulates the attachment/detachment of PC-3 cells and we investigated the role of arctiin on MUC-1 expression. METHODS: The effect of arctiin on PC-3 cell growth was examined using an MTT assay method and cell number was calculated by means of a standard regression line. The expressions of MUC-1 and integrins alpha2, alpha5, and beta1 were detected using FACScan flow cytometric analysis. Levels of MUC-1 mRNA were determined using reverse transcriptase PCR (RT-PCR). RESULTS: Treatment of PC-3 cells with arctiin decreased the cell number in a concentration and time-dependent manner in serum-containing condition. Arctiin preferentially induced cell detachment, but did not have anti-proliferation or cytotoxic effects in PC-3 cells. The arctiin-induced effect was inhibited by cycloheximide, indicating that protein synthesis was required. FACScan flow cytometric analysis demonstrated that arctiin increased the expression of the anti-adhesion mucin MUC 1, but did not affect integrin expression in PC-3 cells. The arctiin-induced increase in MUC-1 protein expression was due to up-regulation of mRNA, as revealed by RT-PCR analysis. CONCLUSIONS: Arctiin significantly induces cell detachment and decreases the cell numbers via the up-regulation of MUC-1 mRNA and protein in PC-3 cells. PMID- 15042602 TI - Overexpressing leptin genetic polymorphism (-2548 G/A) is associated with susceptibility to prostate cancer and risk of advanced disease. AB - BACKGROUND: Leptin has been consistently associated with angiogenesis and tumoral growth. A G/A single nucleotide polymorphism (SNP) at the -2548 site in leptin gene (LEP) is associated with overexpression of leptin (A-allele). METHODS: We evaluated DNA samples from 268 (536 alleles) unrelated individuals, 118 healthy controls (HCs) and 150 prostate cancer (PC) patients, for leptin gene (LEP) locus -2548 genotypes. RESULTS: We found an overrepresentation of the A-allele in PC patients and that there is a significantly higher risk for PC among A carriers (OR = 1.60; confidence interval (CI), 1.13-2.28, P = 0.008). Linear trend analysis showed that quantitative increase of A-allele presence was associated with significantly higher risk for PC (P = 0.003) in heterozygous (OR = 2.11; CI, 1.20-3.71) and homozygous (OR = 2.93; CI, 1.27-6.75) genotypes. Furthermore, the AA and AG genotypes represent significantly higher risk (OR = 4.67; CI, 1.69 12.88 and OR = 2.58; CI, 1.19-5.58, respectively) for advanced disease. CONCLUSIONS: According to our results we hypothesize that the polymorphism in LEP gene may be relevant to PC risk and progression, supporting the hypothesis for leptin involvement in cancer ethiopathogenesis. PMID- 15042603 TI - Ca2+ currents in smooth muscle cells isolated from human prostate. AB - BACKGROUND: The objective of the investigation was to demonstrate the presence of voltage-dependent Ca2+ current in human prostate smooth muscle cells and to determine their biophysical characteristics. METHODS: Prostate smooth muscle cells were isolated from biopsy samples of human prostates obtained from prostatectomy specimens or TURP chips. Electrophysiological recordings were made under current- or voltage-clamp using patch-type electrodes. RESULTS: The average resting potential of prostate myocytes was 63 +/- 11 mV and action potentials (APs) could be elicited when K+ currents were blocked. With K-filled electrodes inward current was followed by a large outward component. When K+ currents were blocked a large Ca2+-sensitive inward current was measured. The inward current could be divided into two components, a fraction blocked by 30 microM verapamil and another by 20 microM NiCl2. CONCLUSIONS: Based on the sensitivity to antagonists and holding potential both L-type and T-type Ca2+ channels were identified in human prostate smooth muscle. PMID- 15042604 TI - 1alpha,25-dihydroxyvitamin D3 induced growth inhibition of PC-3 prostate cancer cells requires an active transforming growth factor beta signaling pathway. AB - BACKGROUND: Prostate cancer growth inhibition by 1alpha,25-dihydroxyvitamin D3 (1,25(OH)2D3) is best characterized in the androgen dependent LNCaP cell line, where treatment with this hormone causes cell cycle arrest and apoptosis. 1,25(OH)2D3 also inhibits the growth of PC-3 prostate cancer cells, but not through the induction of G1 arrest or apoptosis. In this study, we have sought to elucidate the mechanism/s involved in PC-3 cell growth inhibition by 1,25(OH)2D3. EXPERIMENTAL METHODS: We determined the effect of transforming growth factor beta (TGFbeta) blocking antibodies on 1,25(OH)2D3 mediated growth inhibition of PC-3 cells. In addition, we also studied the effects of 1,25(OH)2D3 on TGFbeta signaling and receptor expression. Finally, we assessed the role of TGFbeta signaling in the induction of the growth inhibitory protein, insulin like growth factor binding protein 3 (IGFBP-3), by 1,25(OH)2D3. RESULTS: We find that 1,25(OH)2D3 action in PC-3 cells is mediated through at least two distinct pathways, the TGFbeta pathway and the IGFBP-3 pathway. We show that 1,25(OH)2D3 treatment elevates TGFbeta production and signaling, as well as receptor levels, in PC-3 cells. Further, using a blocking antibody against TGFbeta substantially reduces 1,25(OH)2D3 mediated growth inhibition without affecting IGFBP-3 induction, suggesting that IGFBP-3, alone, is insufficient to inhibit the growth of PC-3 cells. PMID- 15042605 TI - Imaging of bioluminescent LNCaP-luc-M6 tumors: a new animal model for the study of metastatic human prostate cancer. AB - BACKGROUND: Animal experiments examining hormone-sensitive metastatic prostate cancer using the human LNCaP cell line have been limited to endpoint analyses. To permit longitudinal studies, we generated a luciferase-expressing cell line and used bioluminescent imaging (BLI) to non-invasively monitor the in vivo growth of primary LNCaP tumors and metastasis. METHODS: LNCaP.FGC cells were transfected to constitutively express firefly luciferase. LNCaP-luc-M6 cells were tested for bioluminescent signal intensity and hormone responsiveness in vitro. The cells were implanted in subcutaneous and orthotopic sites in SCID-bg mice and imaged over time. RESULTS: The LNCaP-luc-M6 cells formed subcutaneous and orthotopic tumors in SCID-bg mice, and nearly all tumor-bearing animals developed pulmonary metastases. Early detection and temporal growth of primary tumors and metastatic lesions was successfully monitored by BLI. CONCLUSIONS: The LNCaP-luc-M6 cell line is a bioluminescent, hormone-sensitive prostate cancer cell line applicable for BLI studies to non-invasively monitor subcutaneous and orthotopic prostate tumor growth and metastasis in vivo. PMID- 15042606 TI - Serum osteoprotegerin (OPG) levels are associated with disease progression and response to androgen ablation in patients with prostate cancer. AB - BACKGROUND: Osteoprotegerin (OPG) is a tumour and/or bone derived factor that may protect tumour cells from apoptosis. In this study, we have measured serum OPG levels in untreated prostate cancer patients with advanced prostate cancer compared to patients with organ confined disease and in treated patients receiving androgen ablation. METHODS: Serum OPG levels were measured by ELISA in samples collected from 104 patients with either newly diagnosed (n = 59) or advanced prostate cancer treated by androgen ablation (n = 45) and compared with levels in serum from patients with benign prostatic hyperplasia (BPH) (n = 10) and young healthy men (n = 10). RESULTS: Untreated patients with locally advanced disease had significantly higher OPG levels than those with organ confined disease. Patients with advanced disease responding to androgen ablation (serum PSA < 1 ng/ml) had serum OPG levels that were significantly lower than those with clinically progressing disease (PSA > 10 ng/ml). OPG levels in the latter were not significantly different from levels in patients with early signs of biochemical progression (PSA >1 but <10 ng/ml). CONCLUSIONS: OPG is a potential new marker, which is elevated in the serum of patients with advanced prostate cancer and may be an indicator of early disease progression. PMID- 15042607 TI - Screening decreases prostate cancer mortality: 11-year follow-up of the 1988 Quebec prospective randomized controlled trial. AB - PURPOSE: This clinical trial is aimed at evaluating the impact of prostate cancer screening on cancer-specific mortality. SUBJECTS AND METHODS: Forty-six thousand four hundred and eighty-six (46,486) men aged 45-80 years registered in the electoral roll of the Quebec city area were randomized in 1988 between screening and no screening. Screening included measurement of serum prostatic specific antigen (PSA) using 3.0 ng/ml as upper limit of normal and digital rectal examination (DRE) at first visit. At follow-up visits, serum PSA only was used. RESULTS: Seventy-four (74) deaths from prostate cancer occurred in the 14,231 unscreened controls while 10 deaths were observed in the screened group of 7,348 men during the first 11 years following randomization. Median follow-up of screened men was 7.93 years. A Cox proportional hazards model of the age at death from prostate cancer shows a 62% reduction (P < 0.002, Fisher's exact test) of cause-specific mortality in the screened men (P = 0.005). These results are in agreement with the continuous decrease of prostate cancer mortality observed in North America. PMID- 15042608 TI - Combination therapy with vitamins C plus E inhibits survivin and human prostate cancer cell growth. AB - BACKGROUND: We postulated that combinations of C and E vitamins modulate the antioxidant network and blocks survivin gene expression in androgen-responsive and non-responsive human prostate cancer cell (HPCC) lines. METHODS: ALVA-101 and DU-145 cell growth and apoptosis were estimated using the Cell Titer 96 AQ and cell death detection ELISA. Reverse transcription-polymerase chain reaction, Western blot and electrophoretic mobility shift assay were used to quantify survivin mRNA, protein, nuclear factor kappa B (NF-kappaB), and activator protein 1 (AP-1). RESULTS: All the tested combinations of vitamins C and E (25-100 microM, 72 hr) reduced cell growth (4-83%). Vitamin C enhanced the growth suppressive effect of vitamin E. Apoptosis was enhanced (25-45%) (vitamins = 100 microM, 24 hr). Survivin mRNA was decreased (26-29%) (vitamins = 250 microM, 24 hr), and survivin protein was decreased (>90%) (vitamins = 100 microM, 72 hr). NF kappaB and AP-1 activities were increased (vitamins = 100 microM, 24 hr). CONCLUSIONS: The combinations of vitamins C and E are potent inducers of apoptosis in HPCC and suppressers of surviving, an antiapoptotic factor. PMID- 15042609 TI - Expression of serotonin receptors and role of serotonin in human prostate cancer tissue and cell lines. AB - BACKGROUND: Increase in the number of serotonin (5-HT) releasing neuroendocrine (NE) cells has been shown to be correlated with tumor progression, loss of androgen dependence, and poor prognosis. Serotonin is a well-known mitogen which mediates a wide variety of physiological effects via multiple receptors, of which receptor subtype 1 (5-HTR1) has been identified in prostate cancer (PC) cell lines. Recently, 5-HT has been found to show growth-promoting activity and to be functionally related to oncogenes. MATERIALS AND METHODS: Localization, protein content, and mRNA expression of 5-HTR subtype 1A, 1B, and 1D was studied in prostatic tissue (35 patients), metastases, PC cell lines, a benign prostatic stromal cell line (human prostate cell preparation (hPCP)), and xenografts of PC 3 cells by immunohistochemistry (IHC), Western blotting, and RT-PCR, respectively. The growth-inhibition effect of a 5-HT1A antagonist (NAN-190) on PC cell lines was studied using a bromodeoxyuridine (BrdU) assay. RESULTS: A strong immunoreaction of 5-HTR1A and 1B was demonstrated in high-grade tumor cells (35/35) and a small number of BPH cells, whereas 5-HTR1D was confined to vascular endothelial cells. 5-HTR1A was also demonstrated in PC cells metastasized to lymph node and bone, PC-3, DU145, LNCaP, and in xenografts of PC-3 cells and hPCP. Western blot analysis gave strong bands from PC tissue extracts compared to BPH tissue. Using RT-PCR, 5-HTR1A mRNA was demonstrated in all PC cell lines. An antagonist of 5-HTR1A (NAN-190) inhibited the growth of PC-3, DU145, and LNCaP cells but not of hPCP cells. CONCLUSIONS: This is the first study demonstrating an overexpression of 5-HTR subtypes 1A and 1B in PC cells, especially in high grade tumors. Moreover, 5-HT stimulates proliferation of PC cells and 5-HTR1A antagonists inhibit proliferation. Thus, we propose that 5-HT has an important role in tumor progression, especially in the androgen-independent state of the disease. The design of specific antagonists for this type of receptor might be useful for the growth control of androgen-independent tumors. PMID- 15042610 TI - YB-1 is upregulated during prostate cancer tumor progression and increases P glycoprotein activity. AB - BACKGROUND: Currently, the main obstacle to curing advanced prostate cancer is development of androgen independence (AI), where malignant cells acquire the ability to survive in the absence of androgens. Our initial experimental approach used cDNA microarrays to characterize changes in gene expression in the LNCaP human prostate tumor model during progression to AI. The transcription factor Y box binding protein (YB-1) was shown to be one of the genes upregulated. We focused on increased YB-1 expression during progression in clinical specimens, and further examined one of its downstream targets, P-glycoprotein (P-gp). METHODS: Northern blot analysis was performed on LNCaP tumor series, as well as immunohistochemical analyses of human prostate cancer tissue samples. YB-1 was transiently transfected and transport analysis were performed to analyze P-gp efflux activity. RESULTS: YB-1 expression is markedly increased during benign to malignant transformation and further following androgen ablation. In addition, increased YB-1 expression after castration in the LNCaP model is linked to upregulation of P-gp. We demonstrate that YB-1 upregulates P-gp activity resulting in a 40% intracellular decrease in the P-gp substrate vinblastine. We have also found that P-gp increases the efflux of the endogenous androgen, dihydrotestosterone (DHT), from prostate cells and leads to decreased androgen regulated gene expression. CONCLUSIONS: We hypothesize that early in prostate cancer progression, increased expression of YB-1 may increase P-gp activity which may in turn lower androgen levels in the prostate tumor cells. Suppression of androgen levels may activate cell survival pathways and lead to an adaptive survival advantage of androgen independent prostate cancer cells following androgen ablation therapy. PMID- 15042611 TI - Androgen responsiveness of Renilla luciferase reporter vectors is promoter, transgene, and cell line dependent. AB - Renilla based reporters are frequently used as transfection controls for luciferase transcriptional reporter assays. However, recent evidence suggests that a commonly used reporter (HSV-thymidine kinase driven Renilla) is responsive to androgen receptor (AR) and glucocorticoid receptors in the presence of the cognate ligands, dihydrotestosterone (DHT) and dexamethasone (DEX), respectively [1]. We further validate this important technical difficulty by illustrating that in LNCaP prostate cancer cells, spurious Renilla luciferase activity is a function of (a) the promoter driving Renilla expression, (b) the presence of co transfected transgenes, and (c) the androgen responsiveness of the cell line used. Using inhibitors of transcription and translation we showed that transcript interference or translational modulation is not a major means by which androgens affect Renilla luciferase activity. As luciferase reporter assays are a frequent means of studying transcriptional co-regulation in the highly androgen dependent LNCaP cell line, our data serves as a cautionary note that alternative normalization techniques should be employed to avoid misinterpretation of data. PMID- 15042612 TI - Bone microenvironment-related growth factors modulate differentially the anticancer actions of zoledronic acid and doxorubicin on PC-3 prostate cancer cells. AB - OBJECTIVES: We analyzed the actions of zoledronic acid (10-250 microM) and doxorubicin (10-250 nM) on PC-3 prostate cancer cells using both continuous (48 96 hr) and pulsatile exposures (15 min/day for up to three consecutive days). RESULTS: The proliferation of PC-3 cells was inhibited by either continuous or pulsatile exposures of zoledronic acid in a dose-dependent manner. In contrast, pulsatile exposures of doxorubicin failed to inhibit the growth of PC-3 cells. In addition, the inhibition of PC-3 cells by zoledronic acid was partially neutralized by exogenous administration of geranylgeranyl pyrophosphate (GGPP), however, not by farnesyl pyrophosphate (FPP). Furthermore, exogenous administration of transforming growth factor beta 1 (TGF-beta1), interleukin 6 (IL-6), basic fibroblast growth factor (bFGF), and more potently, insulin-like growth factor 1 (IGF-1) inhibited the doxorubicin-induced apoptosis of PC-3 cells. Under identical experimental conditions, these growth factors failed to alter the cytotoxicity of PC-3 cells induced by zoledronic acid. CONCLUSIONS: These data suggest that (i) repetitive and pulsatile (15 min/day) exposure to zoledronic acid inhibited the growth of PC-3 cells, (ii) this anticancer action of zoledronic acid was partially mediated by the attenuation of GGPP production, and (iii) bone microenvironment-related growth factors do not alter the anticancer actions of zoledronic acid on PC-3 cells. PMID- 15042613 TI - Analysis of the macrophage scavenger receptor 1 gene in Swedish hereditary and sporadic prostate cancer. AB - BACKGROUND: The macrophage scavenger receptor 1 (MSR1) gene on chromosome 8p22 was recently reported as a candidate gene for hereditary prostate cancer (HPC). Here, we further elucidate the role of MSR1 in both Swedish families with HPC and in a cohort of unselected prostate cancer. METHODS: DNA samples from 83 Swedish HPC families and 215 unselected population based cases of prostate cancer as well as 425 age-matched controls were genotyped. RESULTS: A total of 18 variants were identified, including 2 exonic, 7 intronic changes, and 9 changes in the 5'- or 3'-uncoding region. Of the two exonic changes, one previously reported truncation mutation was identified, a R293X nonsense mutation. This mutation was found in 2 of the 83 (2.4%) HPC families. The R293X mutation was found more frequently in men with PC (4.9%) than in unaffected men (2.7%), consistent with previous published results, however our results were not significant (P = 0.16). To additionally test for potential association of common sequence variants and increased risk for the disease, five common polymorphisms (PRO3, INDEL1, IVS5-57, P275A, INDEL7) were genotyped in the group of 215 prostate cancer cases and 425 age-matched controls. No association between any of the five common sequence variants and prostate cancer were found. CONCLUSION: Our results suggest that mutations in MSR1 gene might play a role in prostate cancer susceptibility, particularly the R293X mutation. This study warrants further investigations of the role of MSR1 in prostate cancer etiology. PMID- 15042614 TI - The specific role of isoflavones in reducing prostate cancer risk. AB - AIMS: To evaluate the effectiveness of supplementing a group of early stage prostate cancer patients, with 60 mg of soy isoflavones in producing a change in hormonal and proliferative risk parameters that are implicated in prostate cancer promotion. METHODS: Seventy six eligible prostate cancer patients with a Gleason score of 6 or below, between ages 50 and 80 were admitted and supplemented with soy isoflavones or placebo for a 12 week period and changes in PSA and steroid hormones were analyzed at baseline and post intervention. RESULTS: Fifty-nine patients completed the 12-week intervention. Serum free testosterone was reduced or showed no change in 61% of subjects in the isoflavone group compared to 33% in the placebo group. Serum total PSA decreased or was unchanged in 69% of the subjects in the isoflavone treated group compared to 55% in the placebo group. However, we did not see an increase in SHBG levels. Nineteen percent of subjects receiving soy isoflavones reduced total PSA by two points or more during the intervention period. CONCLUSIONS: These data suggest that supplementing early stage prostate cancer patients with soy isoflavones, even in a study of short duration, altered surrogate markers of proliferation such as serum PSA and free testosterone in a larger number of subjects in the isoflavone supplemented group than the group receiving placebo. The study establishes the need to explore further the effects of prolonged and consistent soy consumption, which could potentially delay onset of histologic disease in this patient population. PMID- 15042615 TI - Canine prostate stimulates osteoblast function using the endothelin receptors. AB - BACKGROUND: Bone metastases are common in humans and dogs with late-stage prostate cancer. A unique feature of prostate cancer metastases is new bone formation at metastatic sites ("osteoblastic metastases"). Many carcinomas that metastasize to bone cause bone destruction, not new bone formation. The mechanisms by which prostate cancer induces bone formation at sites of bone metastasis are not well understood. We hypothesized that stimulation of osteoblasts by prostate tissue at metastatic sites was due to the paracrine actions of growth factors produced by prostate epithelial cells. METHODS: We have previously shown that normal canine prostate tissue induced new bone formation when implanted adjacent to the calvarium of nude mice. To complement this in vivo model, we developed an in vitro system of prostate-stimulated osteoblast function to investigate mechanisms of prostate-induced new bone formation. RESULTS: We found that treatment of cultured rat calvaria for 24 hr with proteins from normal dog prostate stimulated alkaline phosphatase activity in a dose-dependent manner 4-6 fold compared to controls. Stimulation began approximately 8 hr after treatment, and was diminished after 72 hr. Calvaria treated with homogenates of normal dog salivary gland, kidney, bladder, and muscle did not increase ALP activity. Pretreatment of the calvaria for 1 hr with endothelin antagonists, but not anti-parathyroid hormone-related protein (PTHrP) antibody or indomethacin, abrogated the stimulation of ALP. CONCLUSIONS: Our results indicated that osteoblast activation by canine prostate occurs via an endothelin-dependent mechanism, and that PTHrP or prostaglandin synthase-mediated pathways are likely not involved. This is a reliable, reproducible assay for determining the roles of molecules important in the activation of osteoblasts by the prostate. PMID- 15042616 TI - Osteopontin enhances the cell proliferation induced by the epidermal growth factor in human prostate cancer cells. AB - BACKGROUND: Susceptibility to extracellular matrix and growth factors has been demonstrated to play a critical role in the development of prostate cancer (PCa) metastases. The aim of this study was to elucidate some mechanisms by which stroma controls tumor progression. METHODS: In our study we tested the growth ability of the LNCaP human prostatic cell line in steroid-free culture conditions in response to osteopontin (OPN), a non-collageneous matrix protein, localized in large amounts in the bone. RESULTS: In the LNCaP cell model, OPN stimulates cell proliferation in serum-free medium and colony growth at high dilution but this effect is visible only in presence of epidermal growth factor (EGF). Proliferation induced by OPN is accompanied by a sustained activation of EGF receptor (EGFR) whose phosphorylation is detectable up to 12 hr after treatment in association with EGF. The colocalization of integrin beta1, a ligand of OPN, and of EGFR on the cellular membrane, suggests that the association of these cell surface receptors may be the principal mechanism involved in the long-term activation of the EGFR. CONCLUSIONS: Our data describe a new possible mechanism involved in the establishment of bone metastases which may also account for the formation of androgen-independent cellular clones, frequently responsible of the clinical progression of PCa. PMID- 15042617 TI - Enhanced invasion of hormone refractory prostate cancer cells through hepatocyte growth factor (HGF) induction of urokinase-type plasminogen activator (u-PA). AB - BACKGROUND: Increased expression of the hepatocyte growth factor (HGF) receptor (MET) is associated with high-grade prostatic adenocarcinoma and metastasis. However, the mechanism through which MET signaling contributes to prostate cancer (CaP) metastasis remains unclear. METHODS: Human PC-3 CaP cells and in vivo selected, isogeneic variant cells of increasing metastatic potential (PC-3M, PC 3M-Pro4, and PC-3M-LN4) were used to investigate the effect of HGF on CaP cell growth, protease production, and invasion. Cell-free urokinase-type plasminogen activator (u-PA) expression and function following HGF treatment were analyzed by Western blot, ELISA, and casein/plasminogen zymography. In vitro invasion stimulated by HGF was measured using Matrigel-coated invasion chambers. RESULTS: Both mRNA and functional protein for MET were detected in each of the CaP cell lines. HGF treatment (0-40 ng/ml) weakly increase proliferation, however, HGF induced soluble u-PA protein and activity 3-fold in the metastatic variant cells. HGF significantly stimulated the invasion of highly metastatic PC-3M-LN4 cells through Matrigel and treatment with specific urokinase receptor inhibitors diminished the HGF-stimulated invasion in a dose-dependent manner. CONCLUSIONS: These results demonstrate the biological significance of u-PA up-regulation in response to HGF in highly metastatic hormone refractory CaP cells. PMID- 15042618 TI - Upregulation and nuclear recruitment of HDAC1 in hormone refractory prostate cancer. AB - BACKGROUND: Histone deacetylase 1 (HDAC1) is a co-repressor involved in differentiation and proliferation control. It is upregulated in malignant compared to benign tissue, and targets a number of transcription factors including p53. METHODS: By immunohistochemistry, HDAC1 protein expression was investigated in human prostate specimens and the CWR22 mouse xenograft model. Flow cytometry and deconvolution immunofluorescence were also performed. RESULTS: HDAC1 was upregulated in pre-malignant and malignant lesions, with the highest increase in expression in hormone refractory (HR) cancer. Using the CWR22 xenograft model we showed androgen dependent regulation of HDAC1. HDAC1 overexpression led to a significant increase in proliferation and a shift towards the undifferentiated cytokeratin (CK) profile in a PC3M derivative clone constitutively expressing HDAC1. CONCLUSION: This study underlines the importance of HDAC1 in cell proliferation and the development of prostate cancer (CaP) and proposes a mechanism for HDAC1 nuclear recruitment. HDAC1 may constitute a crucial therapeutic target particularly in the most lethal phase of androgen independence. PMID- 15042619 TI - USF2 is connected to GAAAATATGATA element and associates with androgen receptor dependent transcriptional regulation in prostate. AB - BACKGROUND: We have previously identified a GAAAATATGATA binding site (pros) of a transcription factor involved in prostatic and androgen-dependent gene regulation. We now purified the potential factors interacting with the pros and characterized their co-operation with the androgen receptor (AR). METHODS: Sequence-specific DNA affinity chromatography, mass-spectrometry, electromobility shift assays, supershifts, glutathione-S-transferase pull-downs, and transient transfections. RESULTS: Several proteins bound to the pros site, but only upstream stimulatory factor 2 (USF2) was confirmed to be part of the transcription factor complex. Weak interaction was detected between AR and the transcription factor complex. Physical proximity between the androgen response element (ARE) and the pros was shown to be important for their co-operation. In the presence of pros and androgen, AR achieves its maximal efficiency even at low concentrations. CONCLUSIONS: The protein complex binding to the GAAAATATGATA site does not have a significant independent function, but may interact with AR if GAAAATATGATA is physically close to the ARE and enhances the transactivation function of AR. PMID- 15042620 TI - Murine prostate cancer inhibits both in vivo and in vitro generation of dendritic cells from bone marrow precursors. AB - BACKGROUND: There is increasing evidence to suggest that dendritic cells (DC) are functionally impaired in tumor bearing hosts. However there is little or no data on the effects of murine prostate cancer (CaP) on DC generation from bone marrow precursors. METHODS: Flow cytometry, mixed leukocyte reactions (MLR), and immunohistochemical analyses were used to characterize DC in CaP. RESULTS: DC generated in the presence of CaP cell lines RM1 and the cell line C2 from the transgenic adenocarcinoma of the mouse prostate (TRAMP) mouse in a Transwell system expressed significantly lower levels of DC differentiation markers. This effect was confirmed when TK-neo-transfected RM1 cells were directly added to DC cultures and eliminated 5 days later using gancyclovir (GCV). Furthermore, co incubation of DC with CaP cells resulted in a decrease in the stimulatory capacity of DC to induce T cell proliferation in the MLR assay. These results were further confirmed in vivo in two different murine models of CaP: i) DC generated from mice intrafemorally injected with TK-neo-transfected RM1 cells; and ii) in DC generated from TRAMP mice. CONCLUSIONS: The generation and function of DC are significantly suppressed in the CaP microenvironment in both in vivo and in vitro murine models. PMID- 15042621 TI - Modulation of androgen receptor-dependent transcription by resveratrol and genistein in prostate cancer cells. AB - BACKGROUND: The androgen receptor (AR) is a ligand-activated transcription factor that mediates the biological responses of androgens in the prostate gland. This study focuses on the chemopreventive agents, resveratrol and genistein, on AR mediated transcription in prostate cancer cells. RESULTS: We found that resveratrol and genistein activated AR-driven gene expression at low concentrations, whereas they repressed the AR-dependent reporter gene activity at high concentrations. We determined that resveratrol and genistein induced AR driven gene expression by activating the Raf-MEK-ERK kinase pathway. The ERK1 kinase phosphorylated the AR on multiple sites in vitro, but this phosphorylation event did not contribute to the resveratrol-induced AR transactivation. CONCLUSIONS: In vitro and in vivo studies have indicated that resveratrol and genistein are promising chemopreventive agents. Given the clear evidence that AR pathways are involved in the development and progression of prostate cancer, these data showed that the ability to modulate AR function would contribute the observed chemopreventive activity of resveratrol and genistein. PMID- 15042623 TI - Studies on microwaves in medicine and biology: from snails to humans. AB - This d'Arsonval Medal acceptance presentation highlights several research themes selected from Dr. Lin's published works, focusing on the microwave portion of the nonionizing electromagnetic spectrum. The topics discussed include investigation of microwave effects on the spontaneous action potentials and membrane resistance of isolated snail neurons, effects on the permeability of blood brain barriers in rats, the phenomenon and interaction mechanism for the microwave auditory effect (the hearing of microwave pulses by animals and humans), the development of miniature catheter antennas for microwave interstitial hyperthermia treatment of cancer, the application of transcatheter microwave ablation for treatment of cardiac arrhythmias, and the use of noninvasive wireless technology for sensing of human vital signs and blood pressure pulse waves. The paper concludes with some observations on research and other endeavors in the interdisciplinary field of bioelectromagnetics. PMID- 15042624 TI - An extremely low frequency magnetic field attenuates insulin secretion from the insulinoma cell line, RIN-m. AB - In this study, we investigated the effects of exposure to an extremely low frequency magnetic field (ELFMF) on hormone secretion from an islet derived insulinoma cell line, RIN-m. We stimulated RIN-m cells to secrete insulin under exposure to an ELFMF, using our established system for the exposure of cultured cells to an ELFMF at 5 mT and 60 Hz, or under sham exposure conditions for 1 h and observed the effects. In the presence of a depolarizing concentration of potassium (45 mM KCl), exposure to ELFMF significantly attenuated insulin release from RIN-m cells, compared to sham exposed cells. Treatment with nifedipine reduced the difference in insulin secretion between cells exposed to an ELFMF and sham exposed cells. The expression of mRNA encoding synaptosomal associated protein of 25 kDa (SNAP-25) and synaptotagmin 1, which play a role in exocytosis in hormone secretion and influx of calcium ions, decreased with exposure to an ELFMF in the presence of 45 mM KCl. These results suggest that exposure to ELFMF attenuates insulin secretion from RIN-m cells by affecting calcium influx through calcium channels. PMID- 15042625 TI - Human head exposure to a 37 Hz electromagnetic field: effects on blood pressure, somatosensory perception, and related parameters. AB - Previous studies have shown that exposure to an electromagnetic field (EMF) of 37 Hz at a flux density of 80 microT peak enhances nociceptive sensitivity in mice. Here we examined the effects on pain sensitivity and some indexes of cardiovascular regulation mechanisms in humans by measuring electrical cutaneous thresholds, arterial blood pressure, heart rate and its variability, and stress hormones. Pain and tolerance thresholds remained unchanged after sham exposure but significantly decreased after electromagnetic exposure. Systolic blood pressure was significantly higher during electromagnetic exposure and heart rate significantly decreased, both during sham and electromagnetic exposure, while the high frequency (150-400 mHz) component of heart rate variability, which is an index of parasympathetic activity, increased as expected during sham exposure but remained unchanged during electromagnetic exposure. Cortisol significantly decreased during sham exposure only. These results show that exposure to an EMF of 37 Hz also alters pain sensitivity in humans and suggest that these effects may be associated with abnormalities in cardiovascular regulation. PMID- 15042626 TI - Response of cyclic AMP by DMSO differentiated HL-60 cells exposed to electric interferential current after prestimulation. AB - The action of interferential current (IFC) upon intracellular content of cyclic adenosine monophosphate (cAMP) after prestimulation with the chemotactic peptide N-formyl-methionyl-leucyl-phenylalanine (fMLP) has been investigated. Human promyelocytes (HL-60) differentiated to granulocytes by dimethylsulphoxide (DMSO) have been treated with different concentrations of fMLP. This enhances their cAMP content. The half maximal effective concentration (EC50) was about 4 nM. Exposure to IFC with modulation frequencies of 35 and 125 Hz (5 min, 250 microA/cm2) after prestimulation with various concentrations of fMLP had no enhancing effect at low or high concentrations of fMLP. In contrast, at medium concentrations in the range of about 100 pM fMLP, a significant enhancement of cAMP could be observed. This synergistic effect of fMLP and IFC has been examined in detail by varying the modulation frequency, current density, and exposure time. Enhancement of cAMP content could be observed at certain modulation frequencies and exposure times suggesting a window effect, whereas for the current density in the range measured (8.5 microA/cm2-2.5 mA/cm2) a constant enhancement could be observed. The synergistic effect of fMLP and IFC could only be observed in the treatment sequence of fMLP followed by IFC; an inverse sequence had no effect on the cAMP content. . PMID- 15042627 TI - Modeling AC current conduction through a human tooth. AB - Impedance between the electrode inserted in a root canal of a human tooth and the outer electrode placed on the oral mucosa serves as a measure of the root canal length, a vital parameter necessary for efficient endodontic procedure in dentistry. For better understanding of current conduction through the tooth, the impedance has been measured on extracted teeth (in vitro) and further used to develop corresponding electrical lumped element models. For modeling the metal/solution interface and complex structure of the tooth, Fricke's constant phase elements are employed. More detailed insight into current conduction is given by numerical simulation. Numerical simulation demonstrates the influence on the impedance of several important parameters, such as dentin conductance, canal preparation, and solution conductance. PMID- 15042628 TI - Resting EEG is affected by exposure to a pulsed ELF magnetic field. AB - An increasing number of reports have demonstrated a significant effect of extremely low frequency magnetic fields (ELF MFs) on aspects of animal and human behavior. Recent studies suggest that exposure to ELF MFs affects human brain electrical activity as measured by electroencephalography (EEG), specifically within the alpha frequency (8-13 Hz). Here we report that exposure to a pulsed ELF MF with most power at frequencies between 0 and 500 Hz, known to affect aspects of analgesia and standing balance, also affects the human EEG. Twenty subjects (10 males; 10 females) received both a magnetic field (MF) and a sham session in a counterbalanced design for 15 min. Analysis of variance (ANOVA) revealed that alpha activity was significantly higher over the occipital electrodes (O1, Oz, O2) [F(1,16) = 6.858; P =.019, eta2 = 0.30] and marginally higher over the parietal electrodes (P3, Pz, P4) [F(1,16) = 4.251; P =.056, eta2 = 0.21] post MF exposure. This enhancement of alpha activity was transient, as it marginally decreased over occipital [F(1,16) = 4.417; P =.052; eta2 = 0.216] and parietal electrodes [F(1,16) = 4.244; P =.056; eta2 = 0.21] approximately 7 min after MF exposure compared to the sham exposure. Significantly higher occipital alpha activity is consistent with other experiments examining EEG responses to ELF MFs and ELF modulated radiofrequency fields associated with mobile phones. Hence, we suggest that this result may be a nonspecific physiological response to the pulsed MFs. PMID- 15042629 TI - 60 Hz electric field upregulates cytosolic Ca2+ level in mouse splenocytes stimulated by lectin. AB - The effect of a 60 Hz electric field (EF) on alteration of cytosolic free Ca2+ level ([Ca2+]c) was examined in mouse splenocytes stimulated by lectins, namely concanavalin A (ConA) or phytohemagglutinin. In order to understand the role of EF on alterations in [Ca2+]c and to determine whether EF exposure increased cell mortality the splenocytes were cultured under the 60 Hz EFs producing current densities of 6 or 60 microA/cm2 for 30 min or 24 h. Cell mortality was less than 2% in experimental all conditions. [Ca2+]c in the splenocyte was not changed by the 6 microA/cm2 exposure alone, while a lectin-induced [Ca2+]c elevation in the EF exposed cells was significantly higher than that of the sham exposed cells (P <.05: ANOVA, P <.05: paired t-test). Moreover, the enhanced increase of [Ca2+]c in the EF exposed, lectin stimulated cells was only observed in the presence of extracellular Ca2+. The EF dependent upregulation of [Ca2+]c persisted after EF exposure (P <.05: paired t-test). The results clearly indicate that Ca2+ influx across the plasma membrane is responsible for the enhanced increase of [Ca2+]c in the EF exposed, lectin stimulated cells and that EF has persistent effect on the cells. Although the precise mechanisms of the EF dependent upregulation of [Ca2+]c is not fully elucidated, the present results demonstrate that the 60 Hz EF (6 microA/cm2) affects [Ca2+]c during cell activation via a Ca2+ influx pathway induced by lectin stimulation. PMID- 15042630 TI - Static magnetic field influence on rat brain function detected by heart rate monitoring. AB - The aim of the present study was to identify the effects of a static magnetic field (SMF) on rat brain structures that control autonomic functions, specifically heart rate and heart rhythmicity. The experiments were carried out on 44 male Wistar rats under ketamine-xylazine anesthesia. SMF was induced using samarium-cobalt fused magnets (20 x 20 x 10 mm in size) placed bitemporally. Magnetic induction intensity was 100 mT on the surface of the head. Duration of magnetic field application was 15 min. An electrocardiogram was recorded from limb lead II, and both heart rate (average duration of cardiac cycles) and heart rhythmicity were analyzed before and after SMF application. SMF evoked changes in both heart rate and rhythm in 80% of the animals; the predominant effects were bradycardia and disappearance of respiratory sinus arrhythmia. However, the effectiveness of SMF in large measure depends on both functional peculiarities and functional activities of brain autonomic centers. PMID- 15042631 TI - Bone morphogenetic protein expression in newborn rat kidneys after prenatal exposure to radiofrequency radiation. AB - Effects of nonthermal radiofrequency radiation (RFR) of the global system of mobile communication (GSM) cellular phones have been as yet mostly studied at the molecular level in the context of cellular stress and proliferation, as well as neurotransmitter production and localization. In this study, a simulation model was designed for the exposure of pregnant rats to pulsed GSM-like RFR (9.4 GHz), based on the different resonant frequencies of man and rat. The power density applied was 5 microW/cm2, in order to avoid thermal electromagnetic effects as much as possible. Pregnant rats were exposed to RFR during days 1-3 postcoitum (p.c.) (embryogenesis, pre-implantation) and days 4-7 p.c. (early organogenesis, peri-implantation). Relative expression and localization of bone morphogenetic proteins (BMP) and their receptors (BMPR), members of a molecular family currently considered as major endocrine and autocrine morphogens and known to be involved in renal development, were investigated in newborn kidneys from RFR exposed and sham irradiated (control) rats. Semi-quantitative duplex RT-PCR for BMP-4, -7, BMPR-IA, -IB, and -II showed increased BMP-4 and BMPR-IA, and decreased BMPR-II relative expression in newborn kidneys. These changes were statistically significant for BMP-4, BMPR-IA, and -II after exposure on days 1-3 p.c. (P <.001 each), and for BMP-4 and BMPR-IA after exposure on days 4-7 p.c. (P <.001 and P =.005, respectively). Immunohistochemistry and in situ hybridization (ISH) showed aberrant expression and localization of these molecules at the histological level. Our findings suggest that GSM-like RFR interferes with gene expression during early gestation and results in aberrations of BMP expression in the newborn. These molecular changes do not appear to affect renal organogenesis and may reflect a delay in the development of this organ. The differences of relative BMP expression after different time periods of exposure indicate the importance of timing for GSM-like RFR effects on embryonic development. PMID- 15042632 TI - Influence of anesthesia on ocular effects and temperature in rabbit eyes exposed to microwaves. AB - To investigate the effect of systemic anesthesia on ocular effects and temperature in rabbit eyes exposed to microwaves, one eye each of 43 male pigmented rabbits (Dutch, 1.8-2.2 kg) was exposed at 2.45 GHz for 60-20 min (300 mW/cm2; 108 W/kg), either under anesthesia (ketamine hydrochloride (5 mg/kg) + xylazine (0.23 mg/kg)) or without anesthesia. Changes in the anterior segment were evaluated by image analysis utilizing a Scheimpflug camera, specular microscopy, and a laser flare cell meter. Temperatures within the eye were measured during microwave exposure by a Fluoroptic thermometer. The exposed eyes showed miosis, conjunctival congestion, corneal edema, and an increase in the light scattering of the anterior shallow cortex in the pupillary area of the lens. The group under systemic anesthesia showed much stronger symptoms than those treated without anesthesia. All of the anterior ocular changes disappeared within a week. The highest temperature during exposure was in the vitreous, followed by the anterior chamber, and the retrobulbar cavity of the orbit. The ocular temperatures of the rabbits under systemic anesthesia were 2-9 degrees C higher than those without anesthesia. Body temperature showed an increase of 1 degrees C during the exposure. Acute high intensity microwave exposure temporarily induced anterior segments inflammation and lens changes. The more pronounced ocular effects in the anesthetized rabbits were associated with the significantly higher ocular temperatures in the anesthetized animals. The influence of systemic anesthesia on ocular changes should be considered. PMID- 15042633 TI - Thriving in an academic environment. PMID- 15042634 TI - "I couldn't think that far": infertile women's decision making about multifetal reduction. AB - In this phenomenological study women's experiences regarding their decisions to undergo or forgo multifetal reduction of their higher-order multiple pregnancies were explored. Seven women who had conceived higher-order multiple pregnancies as the result of in vitro fertilization were interviewed. Four participants accepted reduction, whereas three participants declined. Three themes were discerned: (a) the presence of infertility as a barrier to contemplating hyperfertility; (b) multiple-birth pregnancy as yet another form of loss for infertile women; and (c) the lasting effects of having made the decision. PMID- 15042635 TI - Biochemical hypoglycemia in female nurses during clinical shift work. AB - Female nurses in Japan commonly experience symptoms similar to those of hypoglycemia while working. Biochemical hypoglycemia can lead to impaired cognition; thus, it is important to know the prevalence of biochemical hypoglycemia among nurses. Five hundred and sixty-eight female nurses (53% of the target population) in four hospitals completed questionnaires. They determined their blood glucose levels 12 times, at four points during three shifts. Fifty seven nurses (10%) recorded biochemical hypoglycemia (1 year) by using multivariate regression analyses. The cumulative HBeAg loss rates were 29, 44, and 47% at 1, 2, and 3 years of treatment, respectively. Stepwise Cox's regression analyses indicated that pretherapy viral load was a significant factor associated with HBeAg loss (P = 0.0068). The cumulative emergence rates of YMDD mutants were 23% at 1 year, 45% at 2 year, and 47% at 3 year of treatment. Stepwise Cox's regression analyses indicated that patient age and presence or absence of severe acute exacerbation of liver disease were independent significant factors associated with emergence of YMDD mutants (P = 0.018 and 0.048, respectively). For the development of virological breakthrough, patient age, the presence or absence of severe acute exacerbation, and pretherapy viral load were independent significant factors (P = 0.028, 0.043, and 0.044, respectively). Severe acute exacerbation tended to reduce or delay development of biochemical breakthrough. The present study provides important information for the development of more effective and rational long-term lamivudine therapy for HBeAg-positive chronic hepatitis B patients infected exclusively with genotype PMID- 15042642 TI - Genotypic dominance and novel recombinations in HBV genotype B and C co-infected intravenous drug users. AB - Pathogenic differences among hepatitis B virus (HBV) genotypes have been documented. However, the interaction between different HBV genotypes remains unclear. Herein, we chose HBV genotypes B (HBV/B) and C (HBV/C) co-infected intravenous drug users to study this issue. HBV genotype was determined in 40 HBsAg, anti-HCV, and anti-HDV co-positive intravenous drug users by using genotype-specific primers. The distribution of HBV genotype was as follows: HBV genotype B alone in 29 (72.5%); HBV genotype C alone in 4 (10.0%); and mixed HBV genotype B and HBV genotype C in 7 (17.5%). The interaction between HBV genotype B and HBV genotype C within the same individual was further studied in the seven intravenous drug users with HBV genotype B and HBV genotype C co-infection. By direct sequencing of the pre-S region, only HBV genotype B was detected. When 10 21 clones of the pre-S region were propagated from each intravenous drug user and sequenced, most of the clones were HBV genotype B. Novel recombinations between HBV genotype B and HBV genotype C occurred in four clones (M7-5, M1-10, M1-21, and M1-24) from two intravenous drug users (M7 and M1). The recombination breakpoints were estimated at nucleotide 3120-3171 for M7-5, at nucleotide 3060 3191 for M1-10, and at nucleotide 2910-2950 for M1-21 and M1-24 by SimPlot program. The recombination sites of these HBV/pre-S C-B and B-C recombinants may be within the pre-S1 region. The results in this study suggest that HBV/B is the dominant strain in HBV genotypes B and C co-infected intravenous drug users in Taiwan, and recombinations between different HBV genotype are not unusual. The impact of recombination on the evolution of HBV and their clinical significance remains to be studied. PMID- 15042643 TI - African links and hepatitis B virus genotypes in the Republic of Yemen. AB - Hepatitis B virus (HBV) genotypes in three different communities in the Republic of Yemen with and without significant African links were studied. The subjects included indigenous Yemeni blood donors (n = 987), Africans who had settled down in Yemen for several generations (n = 97) and Soqotrans living on an island in the Indian Ocean close to the African coast (n = 99). Phylogenetic analysis of HBV surface region sequences showed the presence of two major genotypes; A and D: sequences from the blood donors were all genotype D (17/17); those from Africans residing in Yemen were predominantly genotype D (7/8) although one sequence was genotype A; whereas a mixture of genotype A and D was found among the Soqotrans (5/8 and 3/8 respectively). Genotype A sequences were associated with higher viral load, but the difference was not statistically significant. Human migration and international travel, may lead to an interaction between HBV genotypes. The Republic of Yemen is at a location where genotypes A and D meet. The results from this study suggest that genotype A is found only in communities with continuing African links and that genotype D remains the dominant genotype in settled populations. More studies are needed to examine possible long-term changes in HBV genotypes in this region. PMID- 15042644 TI - Diversity and origin of hepatitis B virus in Dutch blood donors. AB - Two considerations led us to study the genetic diversity and origin of hepatitis B virus (HBV) in Dutch blood donors. Firstly, an HBV-infected Dutch blood donor was found negative by four assays used commonly for detection of HBV surface antigen (HBsAg). How variable is HBsAg among HBV infected blood donors? Secondly, the WHO recommends universal vaccination against HBV, but north-west European countries limit vaccination to groups at risk of HBV. This policy may reduce hepatitis B among low-risk, unvaccinated persons if HBV strains that infect low risk persons stem from local at-risk groups. Studying the nucleotide sequence of the S-gene of HBV from 63 Dutch blood donors, considerable variation was found. The majority of the donor strains (52/63, 83%) appears closely related to local HBV isolates as present in intravenous drug users, immigrants, and homosexual men. The remaining 11 (17%) HBV strains belong to various non-Western genotypes. This implies that an indigenous Dutch HBV strain (heterosexually transmitted, not associated with intravenous drug abuse, or immigrants) does not exist, and it supports the policy in low endemic countries to limit vaccination to at-risk groups. On the other hand, it must be realised that, after 20 years of vaccination of at-risk groups, HBV still circulates in the at-risk groups and Dutch blood donors acquire the HBV strains involved. PMID- 15042645 TI - Anti-envelope 1 and 2 immune response in chronic hepatitis C patients: effects of hepatitis B virus co-infection and interferon treatment. AB - Antibodies against envelope glycoprotein 1 and 2 (anti-E1/E2) have been suggested to influence HCV replication levels. Hepatitis B virus (HBV) may interfere with hepatitis C virus (HCV) replication. At present there are no data on anti-E1/E2 antibody responses or on the effect of interferon (IFN) treatment in HBV-HCV co infection. Accordingly, we evaluated serum anti-E1/E2 antibodies in 50 patients (median age, 26.5; males, 30) with chronic hepatitis, 38 with HCV and 12 with HBV HCV co-infection, who had undergone alpha-IFN treatment. Before starting IFN, the HCV group showed higher HCV-RNA levels (bDNA assay) than the HBV-HCV group (median 3.75 vs. 0.64 x 10(6) Eq/ml, respectively; P < 0.05). Similarly, the anti E2 levels (EIA assay) were higher in the HCV group than in the HBV-HCV (mean +/- SD, 53.8 +/- 54.58 vs. 24.5 +/- 41.50 U/ml, respectively; P < 0.02), and the prevalence of anti-E2 was also higher in the HCV group (94 vs. 58%, respectively; P < 0.007). No correlation was found between anti-E1/E2 antibodies and the HCV RNA levels. The prevalence of E1/E2 antibodies was similar in the different HCV genotypes. Higher baseline levels of anti-E2 antibodies and a decrease or disappearance of anti-E2 antibodies during IFN were associated with IFN sustained response in both groups, whereas no reduction in the anti-E1/E2 levels was observed in non-responders. The data show that HBV co-infection influences both HCV replication and the anti-E1/E2 antibody production. High pre-treatment levels of anti-E2 antibodies and their decrease or disappearance during interferon treatment are often associated with HCV clearance in sustained responders, irrespective of the HCV genotype. PMID- 15042646 TI - Prevalence of antibodies to hepatitis E virus among apparently healthy humans and pigs in Bali, Indonesia: Identification of a pig infected with a genotype 4 hepatitis E virus. AB - In Indonesia where hepatitis E virus (HEV) is believed to be highly endemic, only three outbreaks of HEV transmission have been documented to date in restricted areas (West Kalimantan and East Java). A total of 1,115 serum samples collected from apparently healthy individuals in Bali, Lombok, and Surabaya in Indonesia in 1996 where epidemic HEV transmissions have never been reported, were tested for IgG class antibodies to HEV (anti-HEV). In Bali, anti-HEV was detected in 20% (54/276) of the tested population, in remarkable contrast with 4% (17/446) in Lombok and 0.5% (2/393) in Surabaya. On the other hand, antibodies to hepatitis A virus were highly prevalent in all three regions (95% in Bali, 90% in Lombok, and 89% in Surabaya). Although the majority of the population in Indonesia is Moslem, Balinese people are mostly Hindu and have a habit of consuming pork. Therefore, serum samples were obtained from the 99 farm pigs in Bali and tested for anti-HEV and HEV RNA. The sera from 71 pigs (72%) were positive for anti-HEV and a 2-month old pig had detectable HEV RNA. The swine HEV isolate recovered from the viremic pig was named SB66-Bali. The SB66-Bali isolate was most closely related to the genotype 4 isolates from China, India, Japan, and Taiwan, but shared only 82.6 90.0% identity in the common 241-412 nucleotides within open reading frame 2 (ORF2). These results indicate that a presumably indigenous HEV strain(s) is circulating in Bali, Indonesia and that HEV infection may occur via zoonosis even in developing countries. PMID- 15042647 TI - Cytomegalovirus infection after orthotopic liver transplantation is restricted by a pre-existing antiviral immune response of the recipient. AB - Infection of the liver by the human cytomegalovirus (HCMV) frequently occurs after orthotopic liver transplantation (OLT). However, the role of viral replication and the inflammatory reaction in the development of HCMV-associated liver dysfunction is unclear. To address this question in vivo, 84 liver biopsy specimens from 74 patients who received an orthotopic liver transplant were investigated by immunohistochemical detection of viral antigens and cell type specific marker proteins. The extent of viral replication was correlated with the HCMV antibody status of donor and recipient. HCMV immediate early antigens were found in 25 of 84 liver tissue sections investigated, hepatocytes being the predominant target cells. Bile duct epithelial cells, endothelial cells, mesenchymal cells and sinusoidal lining cells were also found susceptible to HCMV infection. The detection of viral capsid antigens, nuclear inclusions in infected cells and foci of infected cells were suggestive of permissive infection in these cells. In 25 HCMV-positive liver biopsy specimens, the median extent of HCMV infection was 0.33 (0.02-5.67) infected cells/mm(2) liver tissue. Primary infection of liver transplant recipients (D+/R-) was associated with a significantly higher extent of organ involvement as compared to reinfection or reactivation (D+/R+). In contrast, the extent of inflammatory infiltrates in areas of infected liver cells was higher in tissues of patients with pre-existing immunoreactivity (R+) compared to patients without pre-existing immunoreactivity (R-). In conclusion, these results favour the assumption that the immune response to HCMV is effective in restricting viral spread in the liver. PMID- 15042648 TI - LightCycler consensus PCR for rapid and differential detection of human erythrovirus B19 and V9 isolates. AB - Until recently, B19 virus was considered to be the only human pathogen of the genus erythrovirus. However, other non-B19 virus strains, such as V9, have now been isolated and are thought to cause infections clinically and serologically indistinguishable from those caused by B19 virus. Whereas B19 virus related isolates have a low genetic diversity of only 1-2%, nucleotide disparity of up to 12% was found for the new isolates, suggesting that non-B19 virus isolates may not be detectable using B19 virus specific PCR methods. To overcome this problem, we designed consensus primers and probes to enable the simultaneous detection of both B19 and non-B19 virus and subsequent discrimination of the two lineages by melting temperature (T(m)) analysis. A total of 196 clinical specimens, from 185 patients with a history of or an anamnesis resembling B19 virus infection, were analyzed using the consensus PCR test. Erythrovirus DNA was detected in 37 of these samples and was found to be B19 virus specific in each case, confirming previous results using B19 virus specific PCR. Although no non-B19 virus DNA was detected in any of the clinical samples tested in this study, more extensive studies are warranted. The routine use of erythrovirus consensus PCR in the diagnosis of B19 virus infection should provide valuable information on the epidemiology and clinical role of non-B19 virus isolates; its use in screening would increase the safety of blood products. PMID- 15042649 TI - Characterisation of penciclovir resistant acyclovir sensitive herpes simplex virus type 2 isolated from an AIDS patient. AB - A heterogeneous herpes simplex virus type 2 (HSV-2) population was characterised from an AIDS patient with relapsing genital ulcer. The isolate had an unusual antiviral spectrum, showing resistance to penciclovir and susceptibility to acyclovir. Two viral populations were plaque purified, one resistant and the other susceptible to both antiviral drugs. The resistant clone was deficient in thymidine kinase (TK) activity and a nucleotide substitution, thymine for cytosine, at position 153 was identified in its TK gene. This mutation resulted in an amino acid change, arginine to tryptophan, in the ATP binding site. In the deficient mutant, a loss of virulence was observed in mice. PMID- 15042650 TI - Human papillomavirus type specific DNA and RNA persistence--implications for cervical disease progression and monitoring. AB - In 2000, we monitored the course and persistence of human papillomavirus (HPV) infection in 54 women who were HPV positive and free of any cytological disease using HPV-DNA genotyping with a linear array assay (baseline). The impact of HPV infection on development of cervical cytological abnormality (dyskaryosis) was monitored by repeat HPV genotyping and cytological assessment 2 years later. Detection of mRNA transcripts of known HPV oncogenes E6 and E7 using NASBA methodology and specific molecular beacons for five common HPV types was also performed at both time points. A total of 11/54 (20%) women developed dyskaryosis after 2 years with 31/54 and 23/54 women exhibiting transient and persistent infections respectively, as monitored by DNA genotyping. Women who maintained type-specific persistent HPV infection were significantly more likely to develop dyskaryosis compared to those who exhibited a transient infection (P = 0.001). The presence of HPV mRNA E6/E7 transcripts was less sensitive but more specific for the detection of disease at follow up. Moreover, women who were DNA positive and also positive for mRNA transcripts at baseline were significantly more likely to harbour persistent infection compared to those in whom DNA only was detected at baseline (P = 0.013). This study highlights the importance of detecting persistent type specific HPV infection to identify those women more at risk of developing cervical abnormalities, either by repeated DNA genotyping, or potentially by RNA based techniques that may be more predictive of persistent infection if performed at a single time point. PMID- 15042651 TI - Antigenic and molecular analysis of influenza A (H3N2) virus strains isolated from a localised influenza outbreak in South Africa in 2003. AB - A severe acute institutional influenza outbreak occurred in a police residential college in Pretoria amongst new recruits and staff members at the end of May 2003. The outbreak was characterised by marked illness which affected a total of 648 students, 26 of whom were admitted to hospital. Symptoms included pyrexia, severe headache, and myalgia. The attack rate per dormitory building ranged from 20 to 47%, with an overall attack rate of 34%. Throat swabs and bronchoalveolar lavage specimens were sent to the National Institute for Communicable Diseases (NICD) from 20 patients. All were positive for influenza A by multiplex PCR and/or indirect immunofluorescence, and were further identified as subtype H3N2. Additional specimens from sporadic influenza cases in Johannesburg and surrounding areas were collected through the NICD active viral surveillance programme for respiratory viral testing and were also positive for influenza A H3N2 viruses. Viruses isolated from patients from both the institutional outbreak as well as from sporadic cases were analysed both antigenically and at the molecular level to determine the characteristics of the influenza strain responsible for the epidemic. The results showed clearly that the outbreak was caused by the introduction in 2003 into South Africa of the novel A/Fujian/411/02 like H3N2 influenza strain, which is antigenically distinct from the A/Panama/2007/99 vaccine strain. The rapid spread of these variant viruses to the southern hemisphere indicates that the H3N2 component of the influenza vaccine needs to be updated for the 2004 southern hemisphere winter. PMID- 15042652 TI - Comparative evaluation of measles virus specific TaqMan PCR and conventional PCR using synthetic and natural RNA templates. AB - Comparative evaluation of TaqMan RT-polymerase chain reaction (PCR) methodology developed during this study with the conventional RT-PCR-nested PCR methodology developed earlier, using measles virus RNA templates derived from synthetic and natural sources against a number of primer sets belonging to various regions of the genome, revealed the existence of similar assay thresholds for both methods. An exception to this finding was, however, noted using primer sets of the N and M genes regions with RNA templates extracted from the wild type measles virus strain where the nested PCR method proved to be 10- to 100-fold more sensitive than the end points established with the N gene specific TaqMan RT-PCR method with synthetic RNA templates. These differences were not evident when the same primer sets were evaluated with RNA templates extracted from a brain sample of SSPE patient. These findings indicate that the genetic make up of measles virus strain in any given clinical specimen, in relation to the amplifying primers/probe sequences, can have impact on the overall sensitivity and specificity of the methodology applied. Both methods are equally suitable for the molecular detection of measles virus sequences in clinical specimens, although the TaqMan RT-PCR method may be preferred due to its advantages of contamination control, automation, and real-time product quantitation. PMID- 15042653 TI - Regional outbreak of mumps due to genotype H in Korea in 1999. AB - Serological and virological studies were carried out of a mumps outbreak which occurred in one region, Yoeju County, Southeast of Seoul in Korea from September to December, 1999. Sera from 736 children at 8-13 years of age of patients with mumps and healthy children were tested for mumps-specific antibodies by enzyme immunoassay. The overall IgM positive rate was 7.6% (56/736), compared with 69.8% (514/736) for IgG. Of the 49 children with both IgG and IgM, 32 were also confirmed by both clinical and serological diagnosis. IgM antibodies were detected even in the samples collected up to 3 months after the onset of symptoms. Although 436 children had been vaccinated before the outbreak, 27 (6.2%) were found to be IgM positive, particularly 6 (4.4%) of 136 were positive serologically despite a second-dose vaccinees. Sequence analysis of the small hydrophobic (SH) gene of 4 mumps viruses isolated from 42 saliva specimens revealed that these were related to the genotype H, but distinguishable from European strains. This is the first study on the outbreak due to mumps virus genotype H and provides information to assess the understanding of recent outbreaks of mumps in Korea. PMID- 15042654 TI - Phylogenetic analysis of clinical mumps virus isolates from vaccinated and non vaccinated patients with mumps during an outbreak, Switzerland 1998-2000. AB - During the past decade mumps outbreaks have occurred in several European countries with universal vaccination programs probably due to poor efficacy of the Rubini vaccine strain. However, the evolution of vaccine escape mutants has also been considered. A phylogenetic analysis was undertaken on 69 clinical mumps isolates obtained from 39 vaccinated and 22 non-vaccinated mumps cases (and six cases with unknown vaccination status) during an outbreak in 1998-2000. Two major strain clusters (SWI-H, SWI-C) with two subgroups each (SWI-H1/2, SWI-C1/2) were identified, which belonged to genotypes C and H. No association between viral clusters and vaccination status or a specific vaccine strain (Jeryl-Lynn or Rubini) was found. Cluster SWI-C1 occurred more frequently in the Western part of Switzerland (P < 0.001). Isolates causing complicated disease tended to cluster more frequently with SWI-H1 (P = 0.11). Wild-type strains homologous or similar to the Rubini vaccine strain (isolated in Switzerland in 1974) were no longer circulating. Therefore, there was no evidence for vaccine escape mutants. Strain redistribution may have occurred during the past decades. Continuous monitoring of circulating mumps virus populations is needed. PMID- 15042655 TI - Molecular epidemiology of mumps virus in Japan and proposal of two new genotypes. AB - We isolated 872 strains of mumps virus from naso-pharyngeal secretions in seven different districts of Japan from January 2000 to July 2001. Among them, 57 strains were geno-typed by nucleotide sequencing in part of the hemagglutinin neuraminidase (HN) and small hydrophobic (SH) protein regions. Four different genotypes (B, G, K, and L) of mumps virus were co-circulating in Japan and the distribution of genotypes varied in geographically different districts. Two new clusters designated as genotypes K and L had more than 7% nucleotide variation in the SH gene. Among the 57 strains, 11 were classified as B, 35 as G, three as K, and eight as L, which was mainly isolated in Tokyo. We also examined 104 stains isolated in a clinic in Mie prefecture from 1993 to 2003. Genotype B was the indigenous strain and genotype K was introduced in 1994. Genotypes B and K co circulated in the 1990s and were replaced by genotype G in 2000. There was no significant change in neutralizing test antibody titers against genotypes B, G, K, and L using seven post-vaccination sera with Hoshino strain (genotype B) and these four genotypes had a different antigenicity from genotype A. We should continue to watch on mumps virus molecular epidemiology. PMID- 15042656 TI - Solubility, immunogenicity and physical properties of the nucleocapsid protein of Nipah virus produced in Escherichia coli. AB - The nucleocapsid (N) protein of Nipah virus (NiV) can be produced in three Escherichia coli strains [TOP10, BL21(DE3) and SG935] under the control of trc promoter. However, most of the product existed in the form of insoluble inclusion bodies. There was no improvement in the solubility of the product when this protein was placed under the control of T7 promoter. However, the solubility of the N protein was significantly improved by lowering the growth temperature of E. coli BL21(DE3) cell cultures. Solubility analysis of N- and C-terminally deleted mutants revealed that the full-length N protein has the highest solubility. The soluble N protein could be purified efficiently by sucrose gradient centrifugation and nickel affinity chromatography. Electron microscopic analysis of the purified product revealed that the N protein assembled into herringbone like particles of different lengths. The C-terminal end of the N protein contains the major antigenic region when probed with antisera from humans and pigs infected naturally. PMID- 15042657 TI - High rate and changing molecular epidemiology pattern of norovirus infections in sporadic cases and outbreaks of gastroenteritis in Hong Kong. AB - Noroviruses (Norwalk-like viruses (NLV)) are recognised as major causes of acute gastroenteritis worldwide. Numerous studies had been carried out on the molecular epidemiology of norovirus in outbreaks but relatively few on sporadic cases. In this study, the molecular epidemiology of noroviruses in sporadic and outbreak cases of acute gastroenteritis in Hong Kong was examined over a 12-month period from July 2001 to June 2002. Specimens from three groups of patients were used in this study. Nine hundred ninety-five specimens from patients enrolled in the Acute Diarrhoeal Diseases Surveillance Programme of the Department of Health, Hong Kong Government; 735 clinical specimens from hospital patients with acute gastroenteritis, and 122 specimens from 44 norovirus outbreaks. Ninety-two (9.2%) surveillance specimens were positive for norovirus RNA by reverse transcription polymerase chain reaction (RT-PCR), compared to 123 (16.7%) clinical and 101 (82.8%) outbreak specimens. For the first 6 months of the study period, the predominant strain was the Bristol strain that belongs to genogroup II (GII). In the latter 6 months of the study, genogroup I (GI) and strains belonging to other clusters of GII were seen more commonly. The vast majority of strains belonging to the Bristol virus cluster were closely related to the 95/96-US subset that was associated with pandemic infection from 1995 onwards. This study clearly establishes the importance of norovirus as a cause of sporadic cases of acute gastroenteritis in all age groups in Hong Kong. PMID- 15042658 TI - Quantitation of group A rotavirus by real-time reverse-transcription-polymerase chain reaction: correlation with clinical severity in children in South India. AB - The epidemiology and pathogenesis of rotaviruses are not completely understood, although recent developments in polymerase chain reaction (PCR) techniques now make it possible to quantify the viral load during an infective episode and investigate its relevance to clinical features of the disease. We studied rotavirus-positive stool samples collected from 10 children without symptoms of gastroenteritis and from 81 children with acute gastroenteritis and in whom the clinical severity of disease was recorded. A semi-quantitative real-time reverse transcription (RT)-PCR was used to estimate the rotavirus load and to assess its correlation with the Vesikari score for severity of diarrhoea. There was a significant negative correlation (r = -0.80, P < 0.001) between severity and the PCR cycle at which the PCR amplicons were detectable (crossing point) on the assay, indicating that children with more severe diarrhoea excrete more virus than children with less severe disease. PMID- 15042659 TI - PCR-probe capture hybridization assay and statistical model for SEN virus prevalence estimation. AB - SEN viruses (SENV) are newly discovered blood-borne single-stranded circular DNA viruses that may play a role in liver disease. To date, no serologic assays are available for the detection of SENV antigens or antibodies. We report on a rapid and sensitive molecular assay for the detection of four SENV strains (SENV-A, -C, -D, -H). This method uses PCR with universal primers and microwell capture hybridization with type-specific probes. Cut-off points to define "infected" based on chemiluminescence readings were determined from a statistical mixture model applied to samples from 300 injection drug users (IDUs) in San Francisco. Based on the estimated cut-off points, we examined the prevalence of SENV infection among 232 healthy US blood donors and assessed sensitivity and specificity of the assay in a small validation sample of infected individuals with partial sequence information. PMID- 15042660 TI - Genetic variation in clinical varicella-zoster virus isolates collected in Ireland between 2002 and 2003. AB - Analysis of genetic variation in 16 varicella-zoster virus (VZV) isolates selected at random and circulating in the Irish population between March 2002 and February 2003 was carried out. A 919 bp fragment of the glycoprotein E gene (open reading frame 68) encompassing codon 150, at which a non-synonymous mutation defines the escape mutant VZV-MSP, and including two other epitope regions e1 and c1, was sequenced. No new single nucleotide polymorphisms (SNPs) were detected, indicating stability of these epitopes in clinical isolates of VZV. However, when four informative polymorphic markers consisting of defined regions from genes 1, 21, 50, and 54 were sequenced 14 variable nucleotide positions were identified. Phylogenetic analysis showed the presence of three highly supported clades A, B, and C circulating in the Irish population. Approximately one third (6/16; 37.5%) of the Irish VZV isolates in this study belonged to genotype C, 4/16 (25%) to genotype A, and 4/16 (25%) to genotype B. A smaller number 2/16 (12.5%) belonged to genotype J1. This indicates remarkable heterogeneity in the Irish population given the small sample size. No evidence was found to suggest any of the 16 isolates was a recombinant. These findings have implications for the model of geographic isolation of VZV clades to certain regions as the circulating Irish VZV population appears to comprise approximately equal numbers of each of the main genotypes. This data is inconsistent with a model of strict geographical separation of VZV genotypes and suggests that VZV diversity is more pronounced in certain areas than had been thought previously. PMID- 15042661 TI - Prevalence of viral infection markers by polymerase chain reaction amplification and interferon-alpha measurements among patients undergoing lumbar puncture in an emergency department. AB - Aseptic meningitis is a frequent diagnosis in emergency departments. Nevertheless, viral investigations are not carried out currently and the viral etiology in adult population has not been studied extensively. We conducted a prospective study including all consecutive patients undergoing lumbar puncture during a 15 months period in an adult emergency department. Bloody and purulent cerebrospinal fluid (CSF) were excluded. The main tests undertaken were: CSF genomic amplification by the polymerase chain reaction (PCR) for neurotropic viruses and serum and CSF interferon-alpha (IFN-alpha) measurements. Among 194 patients included, 45 had and 149 did not have aseptic meningitis. Of 45 patients with aseptic meningitis, 10 had alternative non-virological final diagnosis, and 35/45 were presumed to have neurological disorders of viral origin. Patients (27/35) completed virological analysis: 21/27 (78%) had either positive viral PCR (enterovirus: 8 patients, Varicella zoster virus (VZV): 5, Epstein-Barr virus (EBV): 2, herpes simplex virus (HSV): 1, human herpes virus 6: 1) or only raised serum or CSF IFN-alpha (4 patients). Overall, 59% of patients with a positive viral PCR had either CSF or serum raised IFN-alpha. Twentyone patients without meningitis had either positive viral PCR (enterovirus: 3 patients) or only high serum IFN-alpha level (18 patients). In the setting of aseptic meningitis diagnosed in an adult emergency department, viruses are the most common agents encountered, with enterovirus and VZV as the two main etiological agents. Current CSF viral genome amplification and IFN-alpha measurement are informative and could be useful to confirm the viral origin of various neurological disorders, although the sensitivity and specificity of IFN-alpha measurement for the diagnosis of viral infection need further confirmation. PMID- 15042662 TI - Mumps epididymo-orchitis with prolonged detection of virus in semen and the development of anti-sperm antibodies. AB - Epididymo-orchitis is the most common complication of mumps in post-pubertal men. A case of MMR vaccine failure, in whom mumps and mumps-associated unilateral epididymo-orchitis developed, is presented in this article. Mumps virus was isolated from the semen 14 days after onset and mumps RNA was detected in semen for up to 40 days using RT-PCR. Epididymo-orchitis was associated with transient but significant reduction in sperm count and severe abnormalities in sperm morphology. It also led to the appearance of anti-sperm antibodies, which may have potential long-term adverse effects on the patient's fertility. Sequencing of the SH gene of the virus showed this to be a new mumps genotype distinct from the virus circulating currently in the UK. PMID- 15042663 TI - The ongoing evaluation of breast cancer screening. PMID- 15042664 TI - Time-dependent effects on survival in breast carcinoma: results of 20 years of follow-up from the Swedish Two-County Study. AB - BACKGROUND: Tumor size, lymph node status, and histologic grade are reported to be important predictors of survival in the first 5 years after the diagnosis of invasive breast carcinoma. However, to the authors' knowledge, the effect of these factors in the longer term (>10 years after diagnosis) is not yet clear. METHODS: It is now >20 years since the Swedish Two-County Trial of breast carcinoma screening with mammography was instigated and long-term follow-up is now available to December 1998. In the current study, the authors analyzed the effects of tumor size, lymph node status, and tumor grade on survival to death from breast carcinoma using Cox regression and frailty models that allow the baseline hazard and/or effect of a covariate to vary with time. RESULTS: The effects of tumor size, lymph node status, and tumor grade were shown to progressively diminish with time from diagnosis. The Cox regression model with time-varying coefficients and a dampening parameter then was fitted to allow for the attenuation of prognostic effects; tumor size, lymph node status, and tumor grade were all found to be highly significant (P<0.001). CONCLUSIONS: The results of the current study suggest that long-term survival in women with invasive breast carcinoma could be modelled satisfactorily using either frailty models or Cox regression models with time-varying coefficients. The results also suggest that the value of tumor grade, lymph node status, and tumor size at the time of diagnosis have a lasting influence on subsequent survival, albeit attenuated in later years. The long-term effects of these prognostic factors may explain the fact that the impact of mass screening programs on breast carcinoma mortality rates is still apparent many years later. PMID- 15042665 TI - Breast carcinoma diagnosis, treatment, and prognosis before and after the introduction of mass mammographic screening. AB - BACKGROUND: The introduction of breast carcinoma screening leads to early detection and is believed to reduce mortality and increase the proportion of patients for whom breast-conserving surgery is possible. METHODS: In 1992, a population-based mammographic screening program was introduced in the Dutch city of Tilburg and its surroundings; the program achieved total coverage in 1996. The authors examined the effects of this screening program by investigating disease stage, treatment, and survival among women diagnosed with breast carcinoma at a teaching hospital in Tilburg during the periods 1985-1991 and 1992-1999. RESULTS: Between January 1, 1985, and December 31, 1999, 1400 patients were diagnosed with breast carcinoma. Among patients ages 50-69 years, the proportion of TNM Stage I breast carcinoma increased from 24% in 1985-1991 to 45% in 1992-1999 (P<0.001). The proportion of patients age <50 years with invasive breast carcinoma who underwent breast-conserving surgery decreased from 45% to 33% (P=0.011). Among patients ages 50-69 years, the overall survival rate during the period from 1992 to 1999 was significantly greater than the corresponding rate during the period from 1985 to 1991 (P=0.0009). Even after adjustments were made for tumor stage and patient age, a slight reduction in mortality risk was observed in this age group. No difference in stage distribution or prognosis was found among patients age <50 years or among patients age > or =70 years. Of the 168 invasive malignancies found in patients ages 50-69 years between 1997 and 1999, 68 (40%) were detected by the screening program, 47 (28%) were interval malignancies, and 53 (32%) were detected in nonparticipants or in women who did not participate in 1 or more screening rounds. Patients with screen-detected tumors had a much more favorable prognosis than did patients with interval malignancies (P=0.0018) or patients with clinically detected breast carcinoma (P<0.0001). CONCLUSIONS: Between 1992 and 1999, after the introduction of breast carcinoma screening, improved prognosis and more favorable tumor stage were observed among patients ages 50-69 years. Even after the screening program was fully implemented in 1996, the majority of invasive malignancies still were detected between screening rounds or in patients who did not participate in the program. PMID- 15042666 TI - A Ser49Cys variant in the ataxia telangiectasia, mutated, gene that is more common in patients with breast carcinoma compared with population controls. AB - BACKGROUND: Mothers of children who have ataxia telangiectasia have been reported to be at increased risk for development of breast carcinoma. To test whether sequence variants in the ataxia telangiectasia, mutated, gene (ATM) are associated with breast carcinoma, the authors compared the frequency of ATM cDNA sequence changes in patients with breast carcinoma with the corresponding frequency in control patients. METHODS: The authors sequenced ATM cDNA from 91 patients with breast carcinoma and compared the frequencies of sequence changes in these patients with the corresponding frequencies in a control sample of 940 individuals with no history of malignant disease. RESULTS: Thirty-five patients with breast carcinoma had one or more single-base changes in ATM. Three genetic variants were found in at least two patients. These variants resulted in Asp1853Asn, Pro1054Arg, or Ser49Cys amino acid substitutions in the ATM protein. The Ser49Cys variant was more common in patients with breast carcinoma than in the control patients, with respective frequencies of 6.7% (5 of 75 patients) and 1.3% (12 of 940 patients; P=0.006; Fisher two-sided exact test). The subgroup of patients with bilateral breast carcinoma had a Ser49Cys frequency of 11.8% (2 of 17 patients), which again was significantly different from what was observed in the control group (P=0.024; Fisher two-sided exact test). The allele frequencies of the other two variants were not different between case patients and control patients. CONCLUSIONS: Patients with breast carcinoma, particularly those with bilateral disease, were more likely to have a variant in the ATM gene that resulted in a Ser49Cys substitution in the gene product. Additional studies are needed to evaluate the potential functional consequences of the Ser49Cys substitution and confirm the relevance of this variant in the development of breast carcinoma. PMID- 15042667 TI - DNA repair and breast carcinoma susceptibility in women. AB - BACKGROUND: Breast carcinoma is the most common cancer and the second leading cause of cancer-related deaths among women. The disease represents approximately 31% of all cancers in Puerto Rican women. Several DNA repair pathways are involved in preventing carcinogenesis. The current study evaluated the hypothesis that a reduced DNA repair capacity (DRC) is a susceptibility factor for breast carcinoma. METHODS: A retrospective case-control clinical study was performed to compare age-matched DRC in 33 women with histopathologically confirmed breast carcinoma (cases) and 47 cancer-free women (controls). DRC was measured using a host cell reactivation assay with a luciferase reporter gene and then transfected into human peripheral lymphocytes. A questionnaire was used to solicit breast carcinoma risk factors. RESULTS: Women with breast carcinoma had a mean DRC of 5.6% +/- 0.5 standard error of the mean (SEM). Cancer cases had a 36% reduction (P<0.001) in DRC when compared with the control group (DRC=8.7% +/- 0.7 SEM). Younger participants with breast carcinoma were found to have a more significant reduction in DRC when compared with age-matched controls. Family (odds ratio [OR]=4.1), maternal lineage (OR=5.5), and maternal (OR=12.4) history of breast carcinoma were found to be the only statistically significant (P<0.05) risk factors associated with the disease. CONCLUSIONS: The findings supported the hypothesis that a low DRC is a susceptibility factor for breast carcinoma. A 1% decrease in DRC corresponded to a 22% increase in breast carcinoma risk. To the authors' knowledge, the current study was the first to directly determine the DRC of women with breast carcinoma. Because DRC is an independent risk factor for breast carcinoma, the DRC of women may be a useful marker in predicting susceptibility. PMID- 15042668 TI - A prospective study of concurrent cyclophosphamide/methotrexate/5-fluorouracil and reduced-dose radiotherapy in patients with early-stage breast carcinoma. AB - BACKGROUND: Concurrent administration of chemotherapy and radiotherapy has the potential advantage of delaying neither treatment and providing radiation sensitization. However, the optimal approach to concurrent treatment in women with early-stage breast carcinoma remains undefined. We present updated results of a prospective protocol of concurrent cyclophosphamide/methotrexate/5 fluorouracil (CMF) and reduced-dose radiotherapy, focusing on tumor control and patient tolerance. METHODS: One hundred twelve women with AJCC Stage I or Stage II breast carcinoma with 0-3 positive axillary lymph nodes were enrolled in a prospective single-arm study of concurrent CMF and reduced-dose radiotherapy (39.6 gray [Gy] to the whole breast, 16-Gy boost). A high proportion of women had risk factors associated with an increased risk of local disease recurrence, including age <40 (32%), close or positive margins (37%), or lymphatic/vascular invasion (51%). The median follow-up period was 94 months. RESULTS: The 5-year overall survival rate was 94%. By 60 months, 5 patients (4%) experienced local disease recurrence and 19 patients (17%) experienced distant metastasis. There were no isolated regional lymph node recurrences. Local disease recurrence occurred in 1 of 25 patients (4%), 1 of 16 patients (6%), and 3 of 70 patients (4%) with positive, close (<1 mm), and negative margins, respectively. One patient developed acute myelogenous leukemia. An additional patient developed Grade 2 pneumonitis. Cosmetic results were not recorded uniformly for all patients and therefore could not be reliably analyzed. CONCLUSIONS: Concurrent CMF and reduced-dose radiotherapy resulted in a low level of late toxicity and excellent local tumor control, despite the large proportion of patients with substantial risk factors for local disease recurrence. Future studies of concurrent regimens, particularly in patients at high risk of local disease recurrence, are warranted. PMID- 15042669 TI - Change in tumor cellularity of breast carcinoma after neoadjuvant chemotherapy as a variable in the pathologic assessment of response. AB - BACKGROUND: Complete pathologic response of breast carcinoma to neoadjuvant chemotherapy is a well defined outcome that correlates with prolonged survival. Categorization of incomplete response depends on accurate measurement of residual tumor size but is complicated by the variable histopathologic changes that occur within the tumor bed. In the current study, the authors investigated the contribution of assessing tumor cellularity in the pathologic evaluation of response to chemotherapy. METHODS: The slides from diagnostic core needle biopsy and the subsequent matched resection specimens were examined in 240 patients with breast carcinoma: 120 "treated" patients who received neoadjuvant chemotherapy and 120 "control" patients who received primary surgical management within a few weeks of diagnosis. Clinical response and residual tumor size were evaluated in 108 treated patients who completed a clinical trial with paclitaxel and then received combined 5-fluorouracil, doxorubicin, and cyclophosphamide chemotherapy. Tumor cellularity was assessed from hematoxylin and eosin-stained tissue sections as the percentage of tumor area that contained invasive carcinoma. RESULTS: After neoadjuvant chemotherapy, tumor cellularity decreased from a median of 40% in core needle biopsy to 10% in resection specimens (P<0.01; Wilcoxon signed rank test). The cellularity of core needle biopsy (median, 30%) tended to underestimate the cellularity of resection specimens (median, 40%) in the control group (P<0.01). Changes in cellularity varied within each clinical response category, particularly partial response and minor response. The greatest reduction was observed in the cellularity of residual primary tumors that measured < or =1 cm (pathologic T1a [pT1a] and pT1b tumors), but changes in cellularity varied in the pT1, pT2, and pT3 residual tumor categories. The shape of the distribution of tumor size, expressed as the greatest dimension in cm, was similar in the control group and the treatment group (excluding complete pathologic response); however, when residual tumor size and cellularity were combined, the distribution of pathologic response shifted left (toward complete response) with a steep decline, suggesting that many tumors had a large reduction in cellularity but little change in the tumor size. CONCLUSIONS: Cellularity of the tumor mass was reduced significantly by neoadjuvant chemotherapy, and the change varied widely in different categories of clinical response. Although residual tumors measuring < or =1 cm in greatest dimension had the most reduction in tumor cellularity, there was broad variability for all residual tumor groups (pT1-pT3). The frequency distribution of residual tumor size was altered markedly by the inclusion of tumor cellularity, indicating that the product of pathologic size and tumor cellularity may provide more accurate pathologic response information than tumor size alone. PMID- 15042670 TI - An assessment of World Health Organization criteria for severe acute respiratory syndrome in patients with cancer. AB - BACKGROUND: The differential diagnosis of severe acute respiratory syndrome (SARS) in patients with cancer can be challenging. Although diagnostic criteria for SARS have been issued by the World Health Organization (WHO), simple adoption of the established criteria may lead to overdiagnosis in patients with cancer or to an increase in the risk of spreading SARS within cancer hospitals. METHODS: The authors report their experience with the exclusion and quarantine of patients with cancer during the peak of the SARS epidemic in Beijing, China. The patients included 4 males and 7 females with a median age of 66 years (range, 39-73 years). RESULTS: All 11 patients met the WHO diagnostic criteria for probable SARS. Among those 11 patients with probable SARS, only 1 had confirmed SARS; for the other 10 patients, the possibility of SARS infection was ruled out. CONCLUSIONS: Special attention must be paid to patients with cancer who have symptoms similar to those seen in SARS. Although the WHO diagnostic criteria for SARS should be widely accepted, they are not definite or practical in certain populations. PMID- 15042671 TI - X-chromosome loss of heterozygosity frequently occurs in gastrinomas and is correlated with aggressive tumor growth. AB - BACKGROUND: Recent studies have shown that tumor growth, rather than hormone overproduction, is the leading cause of death among patients with gastrinomas and other malignant gastrointestinal endocrine tumors. No patient/laboratory characteristics accurately predict which tumors will exhibit aggressive growth. Furthermore, little is known regarding the molecular pathogenesis of these tumors. X-chromosome loss of heterozygosity (LOH) occurs in some nonendocrine tumors, and its presence can be associated with aggressive growth/decreased survival. Data on X-chromosome LOH in gastrointestinal endocrine tumors are conflicting. Therefore, the purpose of the current study was to determine whether X-chromosome LOH occurred in gastrinomas and, if so, whether it was correlated with tumor growth, tumor behavior, and/or prognosis. METHODS: X chromosome allelotyping was performed using 12 microsatellite markers spaced throughout the chromosome using DNA from leukocytes and microdissected gastrinoma specimens from 16 female patients. The presence of X-chromosome LOH was analyzed for correlations with clinical and laboratory tumor characteristics as well as tumor growth characteristics. RESULTS: Nine gastrinoma specimens (56%) had X-chromosome LOH, ranging from 6% to 23% at the 12 different loci studied. X-chromosome LOH was significantly associated with aggressive postoperative tumor growth, increased primary tumor size, and pancreatic primaries. In 6 tumor specimens, LOH occurred on Xp22.1-22.3 over a 28.4-centimorgan region. CONCLUSIONS: X-chromosome LOH was common in gastrinoma specimens from female patients, and its presence was found to be a potentially useful molecular/genetic prognostic factor for aggressive growth. PMID- 15042672 TI - Interleukin-1 (IL-1alpha and IL-1beta) and its receptors (IL-1RI, IL-1RII, and IL 1Ra) in prostate carcinoma. AB - BACKGROUND: The principal components of the interleukin-1 (IL-1) family are two secreted factors (IL-1alpha and IL-1beta), two transmembrane receptors (IL-1RI [biologically active] and IL-1RII [inert receptor]), and a natural antagonist receptor of IL-1 function (IL-1Ra). Changes in the expression pattern of these IL 1 members have been reported to be related to disease progression. The objective of the current study was to evaluate these changes in prostatic tissue by means of immunohistochemistry and Western blot analysis. METHODS: Immunohistochemical and Western blot analyses were performed in 20 normal samples, 35 samples of benign prostatic hyperplasia (BPH) and 27 samples from patients with prostate carcinoma (PC). RESULTS: In normal prostate samples, immunoreactions to IL-1beta and IL-1RI were positive, whereas there were no immunoreactions observed to IL 1alpha, IL-1RII, or IL-1Ra. In BPH, in addition to immunoreactions to IL-1beta and IL-1RI, immunoreactions to IL-1alpha, IL-1RII, and IL-1Ra were observed in many samples. In samples of PC with low Gleason grade, most tumors had positive immunoreactions to IL-1alpha and IL-1RI. In samples of PC with high Gleason grade, immunoreactions were seen only to IL-1alpha, IL-1RI, and IL-1RII. CONCLUSIONS: The current results suggested that high expression levels of IL 1alpha and IL1-RI in epithelial cells in BPH and PC samples were involved in cell proliferation and that the loss of immunoexpression of IL-1beta and IL-1Ra was a characteristic feature of PC compared with normal prostate samples and BPH. Because this loss is progressive up to a complete absence of immunoexpression in PC of high Gleason grade, the evaluation of IL-1beta and IL-1Ra in PC may be significant in assessing for malignancy. PMID- 15042673 TI - Results of a randomized, population-based study of biennial screening using serum prostate-specific antigen measurement to detect prostate carcinoma. AB - BACKGROUND: The purpose of the current study was to evaluate the effectiveness of a prostate carcinoma screening program in which serum prostate-specific antigen (PSA) levels were measured. METHODS: From a group of 20,000 men born between January 1, 1930, and December 31, 1944, 10,000 men were randomized into a screening group and 10,000 were randomized into a control group. Patients in the screening group were invited to undergo initial PSA testing between 1995 and 1996 and then were invited to receive testing every second year thereafter for 8 years (for a total of 4 PSA tests). Men with PSA levels > or =3 ng/mL (or > or =2.54 ng/mL, in the third and fourth screening rounds) were invited to undergo clinical investigation, which included sextant biopsy of the prostate. By linking to the regional cancer registry, the authors were able to obtain the true and expected incidence rates for the screening and control groups. RESULTS: The screening participation rate was high (73%). A total of 884 malignancies have been detected to date, with 640 having been detected in the screening group. There was an early and marked shift toward more favorable disease stage and grade for malignancies detected on repeat screening. In the fourth screening round, only 2 of 82 detected malignancies were classified as advanced disease. Of the 227 screen detected tumors on which surgery was performed, only 20 (8.8%) had small volume (<0.2 cm3). Forty-three interval malignancies were detected, but only five were accompanied by symptoms. At 8 years, the cumulative disease incidence rate among screening participants was 7.3%, compared with 2.4% in the control arm. The incidence rate observed in the screening population corresponds to the cumulative incidence rate observed in the Swedish male population at age 72 years. CONCLUSIONS: Biennial PSA screening was very successful in diagnosing prostate carcinoma at an early stage, when curative treatment typically is effective. In addition, the results regarding interval malignancies were favorable. Thus, decreased mortality should be observed on long-term follow-up. The lead time associated with screening appears to fall within the range described in earlier studies involving frozen sera (i.e., 5-9 years). PMID- 15042674 TI - Chromophobe renal cell carcinoma: analysis of 61 cases. AB - BACKGROUND: Chromophobe renal cell carcinoma (CRCC) is often associated with a favorable prognosis. However, to the authors' knowledge, only few clinical data are available regarding this variant of tumor. In the current study, the authors report their experience with CRCC over the last 14 years. METHODS: Since 1989, 61 patients have been treated at the study institution for CRCC. Tumor characteristics and patient outcome were analyzed retrospectively. Data were obtained from the patients' medical records. RESULTS: The mean age of the patients was 58 years. Of the 61 tumors, 68.8% were discovered incidentally. The mean tumor size was 6.9 cm. Fifty-seven patients (93.4%) were treated with radical nephrectomy and 4 patients (6.6%) underwent partial nephrectomy. According to the 1997 TNM classification, the pathologic tumor stage was T1 in 65.6% of cases, T2 in 31.1% of cases, and T3a in 3.3% of cases. All tumors were staged as N0M0. Nuclear grade was low (1 or 2) in 88.5% of cases. In no case of CRCC was a sarcomatoid component observed. At a mean follow-up of 49.5 months (range, 5-135 months), no patient had experienced tumor recurrence or disease progression, and none had died of renal carcinoma. CONCLUSIONS: In the authors' experience, CRCC carries an excellent prognosis, possibly due to the high rate of low-stage and low-grade tumors. PMID- 15042675 TI - Metastatic trophoblastic disease after an initial diagnosis of partial hydatidiform mole: genotyping and chromosome in situ hybridization analysis. AB - BACKGROUND: Hydatidiform mole (HM) is classified into partial (PHM) and complete (CHM) subtypes according to histopathologic and genetic criteria. Traditionally, it is believed that PHM carries a better prognosis and rarely develops metastasis. However, making a distinction between PHM and CHM using histologic criteria alone may be difficult. METHODS: The authors used fluorescent microsatellite genotyping following laser-capture microdissection and chromosome in situ hybridization (CISH) to perform a genetic analysis of six patients with histologically diagnosed PHM who subsequently developed metastatic gestational trophoblastic neoplasia. RESULTS: Patients ranged in age from 25 years to 44 years (mean, 33.2 years). The gestational age of the molar pregnancies varied from 6 weeks to 20 weeks. All six patients had pulmonary metastases, with additional liver metastasis in two patients. Among the six patients with histologically diagnosed PHM, it was found that four patients had a diploid karyotype and no maternal alleles; thus, their neoplasms actually were CHM. Maternal genome was detected in the remaining two patients consistent with a biparental origin, and these patients had a triploid karyotype. CISH findings in all patients correlated with the genotyping findings. Triploid HM had maternally derived alleles, whereas diploid HMs were purely androgenetic. CONCLUSIONS: In the current study, which may be the largest series of genetically analyzed metastatic PHMs to date, the difficulty of histologic distinction between PHM and CHM was confirmed. Molecular analysis may help to refine the classification of HM. Although the current findings support the belief that most aggressive trophoblastic diseases are derived from CHM, a small number of PHMs do progress to metastatic disease. Thus, the current study reaffirmed that all patients with HM should be followed closely irrespective of histologic subclassification. PMID- 15042676 TI - Neoadjuvant therapy changes the lymphocyte composition of tumor-draining lymph nodes in cervical carcinoma. AB - BACKGROUND: The objective of the current study was to illustrate the influence of neoadjuvant therapy on the local immune response in patients with cervical carcinoma. METHODS: Uninvolved tumor-draining lymph nodes (TDLN) (n=158 lymph nodes), including internal, external, and common iliac lymph nodes as well as obturator, presacral, and aortic lymph nodes from 15 nontreated (NT) patients, 4 chemotherapy (CT)-treated patients, and 19 chemoradiation (CR)-treated patients, were analyzed for lymphocyte subset distribution and for the proliferative response of T cells to polyclonal activation and interferon-gamma (IFN-gamma) production. Lymphocyte subsets in peripheral blood also were assessed. RESULTS: TDLNs from CR-treated patients contained higher proportions of CD8+ cells and natural killer cells than NT and CT-treated patients (P values ranged from <0.05 to <0.01). TDLNs from CR-treated patients were enriched in activated-type CD4+ cells (HLA-DR+, CD134+, CD62L-, and CD25+ at an intermediate expression level; P values ranged from <0.05 to <0.01) and activated-type CD8+ cells (CD62L-, P<0.001) compared with NT patients. Concomitantly, there was a reduction in the proportion of naive-type CD4+ and CD8+ cells (CD45RA+/CD62L+) (P<0.01 and <0.05, respectively). CR treatment increased the proportion of both CD4+ and CD8+ cells prone to produce IFN-gamma. All TDLNs contained suppressive CD4+ T regulatory (Treg) cells (CD25+ and CD152+ at a high expression level) whose frequency and suppressive activity was not influenced by the treatment. Therapy-induced changes in TDLN were mirrored only in part by respective alterations in peripheral blood. CONCLUSIONS: To our knowledge, the current study is the first to show that neoadjuvant therapy produces an enhancing effect on the immune competency of TDLNs from patients with cervical carcinoma. PMID- 15042677 TI - Comparison of clinical and molecular surveillance in patients with advanced nasopharyngeal carcinoma after primary therapy: the potential role of quantitative analysis of circulating Epstein-Barr virus DNA. AB - BACKGROUND: The importance of detecting recurrence at an early stage in patients with malignant disease is well recognized. Circulating Epstein-Barr virus (EBV) DNA can be detected in patients with nasopharyngeal carcinoma (NPC). The objective of the current study was to assess the effectiveness of plasma EBV DNA monitoring in the early detection of NPC recurrence compared with conventional methods. METHODS: Patients with NPC in two prospective clinical trials who had locoregional recurrences or distant metastases were recruited into the study. Clinical data on these patients were scrutinized for evidence of recurrence. EBV DNA copy numbers in the prospectively collected plasma samples were assayed retrospectively with real-time quantitative polymerase chain reaction analysis. RESULTS: At the time of clinical recurrence, 65% of 26 patients with locoregional recurrences and all but 1 of 28 patients with distant failure had circulating EBV DNA. The difference between the time from completion of treatment to positivity for circulating EBV DNA and the time from completion of treatment to the first observed clinical abnormality was not statistically significant for patients with local recurrence (P=0.84). However, the time to the first detection of circulating EBV DNA was significantly shorter among patients with distant metastases (P<0.0001). The Kaplan-Meier estimated median time to the emergence of plasma EBV DNA was 190 days, with a 95% confidence interval (CI) of 95-300 days, and the median time to the first observed clinical abnormality was 295 days (95% CI, 276-361 days). CONCLUSIONS: Monitoring plasma EBV DNA levels surpassed traditional methods for the early detection of distant failure in patients with NPC. The role of this technique should be evaluated in prospective studies that incorporate complementary advanced imaging technology. PMID- 15042678 TI - Intensive chemotherapy with and without cranial radiation for Burkitt leukemia and lymphoma: final results of Cancer and Leukemia Group B Study 9251. AB - BACKGROUND: The objective of the current study was to evaluate the efficacy of intensive chemotherapy with and without cranial radiation for central nervous system (CNS) prophylaxis in adults with Burkitt leukemia or lymphoma. METHODS: Patients received 18 weeks of therapy. Prophylactic cranial radiation (2400 centigrays) and 12 doses of triple intrathecal chemotherapy were administered to the first cohort of patients. A subsequent cohort received the same therapy, with the exceptions that intrathecal therapy was reduced to six doses and radiotherapy was administered only to high-risk individuals. RESULTS: The median follow-up durations were 6.8 years in Cohort 1 and 4.1 years in Cohort 2. Three occurrences of transverse myelitis, 2 severe neuropathies, 3 cases of aphasia, and 1 case of blindness were documented in the first cohort of 52 patients (Cohort 1). In the subsequent cohort of 40 patients (Cohort 2), none of these occurrences were observed, and patients experienced less neurologic toxicity overall (61% vs. 26%; P=0.001). Responses were similar, and the 3-year event-free survival rate was 0.52 (95% confidence interval, 0.38-0.65) for Cohort 1 and 0.45 (0.29-0.60) for Cohort 2. CONCLUSIONS: Intensive, short-duration chemotherapy with less intensive CNS prophylaxis led to control at this sanctuary site with little neurotoxicity and may be curative for adults with Burkitt leukemia or lymphoma. PMID- 15042679 TI - Phase I and pharmacokinetic study of a low-clearance, unilamellar liposomal formulation of lurtotecan, a topoisomerase 1 inhibitor, in patients with advanced leukemia. AB - BACKGROUND: OSI-211 is a low-clearance, unilamellar liposomal formulation of a water-soluble camptothecin analogue, lurtotecan. OSI-211 has significant activity in severe combined immunodeficient mouse models of human leukemia. METHODS: This study was conducted to define the dose-limiting toxicities (DLT) and pharmacokinetics of OSI-211 in patients with refractory myeloid leukemias. Patients with refractory acute myeloid leukemia (AML), myelodysplastic syndrome (MDS), or chronic myelogenous leukemia in blastic phase (CML-BP) were eligible. OSI-211 was given as an intravenous infusion over 30 minutes daily for 3 days. The starting dose was 1.5 mg/m2 per day (4.5 mg/m2 per course). The dose was escalated by 50% until Grade 2 toxicity was observed and then by 30-35% until the DLT was defined. Serial plasma and urine samples were collected, and drug levels were determined by high-performance liquid chromatography with fluorescence detection. RESULTS: Twenty patients (18 patients [90%] with AML, and 1 patient each [5%] with MDS and CML-BP) were treated. Mucositis and diarrhea were considered to be the DLTs. The maximum tolerated dose was 3.7 mg/m2 per day. Fourteen of 18 evaluable patients (78%) with AML or MDS achieved transient bone marrow aplasia. The mean systemic clearance of lurtotecan in plasma was 0.946 +/- 1.53 L/hour/m2. Urinary recovery of lurtotecan was 6.66% +/- 5.26% (range, 1.05 18.4%). CONCLUSIONS: Liposomal encapsulation of lurtotecan altered its metabolism significantly. There was no evident correlation between exposure, as measured by plasma pharmacokinetics of lurtotecan, and clinical response or toxicities. OSI 211 merits further study in hematologic malignancies. PMID- 15042680 TI - Changes associated with the development of resistance to imatinib (STI571) in two leukemia cell lines expressing p210 Bcr/Abl protein. AB - BACKGROUND: Although various mechanisms have been recognized as being associated with the development of resistance to imatinib mesylate in vitro and in clinical situations, their relative significance and contributions remain poorly understood, as is the sequence of events leading to the selection of the resistant phenotype. Experimental in vitro systems involving well defined cell lines and conditions can be used to some advantage to answer specific questions and to develop in vitro models of imatinib resistance that would reflect its potential heterogeneity. METHODS: Two cell lines, KBM5 and KBM7, which expressed p210 Bcr/Abl and which differed in their inherent sensitivity to imatinib, the number of copies of the BCR/ABL fusion gene, and the activation of apoptotic pathways, were grown in vitro in the presence of increasing concentrations of imatinib. The resistant cells were analyzed for cell cycle progression, apoptotic response after exposure to imatinib, expression of Bcr/Abl, tyrosine kinase activity, and the presence of mutations within the adenosine triphosphate (ATP) coding domain of BCR/ABL. At various levels of resistance, the cells were transferred into drug-free media, and the stability of the resistant phenotype was determined in the absence of the drug. RESULTS: In KBM7 cells, the development of resistance was characterized by loss of apoptotic response to the drug, amplification of BCR/ABL, increased levels of expression of p210 Bcr/Abl, and decreased inhibition of Bcr/Abl tyrosine kinase (TK) activity by imatinib. No mutations within the ATP-binding domain of Bcr/Abl were identified, and resistance remained stable in the absence of the drug. In KBM5 cells, which previously were found to be characterized by the acquisition of a single C-T mutation at ABL nucleotide 944 (T315I) at high levels of resistance, this same mutation was detected at an intermediate level, but not at a low level, of resistance. The response of KBM5 cells to imatinib was characterized by a low level of apoptotic response, a marginal increase in BCR/ABL copy number, a modest increase in p210 expression, and a highly imatinib-resistant Bcr/Abl TK. Partial reversal of resistance was observed in highly resistant KBM5-STI571(R1.0) cells, which continued to display the C-T mutation. In KBM5 cells with an intermediate level of resistance, the T315I mutation was no longer detectable upon their reversal to the sensitive phenotype. CONCLUSIONS: BCR/ABL amplification with subsequent overexpression of Bcr/Abl protein, loss of apoptotic response, or point mutation of the ATP-binding site of BCR/ABL was associated alternatively with the acquisition of the resistant phenotype, supporting the notion that multiple mechanisms are involved in the induction of resistance to imatinib. The initial number of BCR/ABL copies itself was not related directly to the degree of resistance. The reversibility of the resistance may be complete, partial, or irreversible, depending on the mechanism(s) involved and the degree of resistance. Both cell lines serve as models for further elucidation of various aspects of imatinib-resistance mechanisms. PMID- 15042681 TI - Hypermethylation of FHIT as a prognostic marker in nonsmall cell lung carcinoma. AB - BACKGROUND: Methylation of CpG islands in the promoter and upstream coding regions has been identified as a mechanism for transcriptional inactivation of tumor suppressor genes. The purpose of the current study was to determine the correlation between the aberrant promoter methylation of multiple genes and survival in patients with nonsmall cell lung carcinoma (NSCLC). METHODS: The methylation status of nine genes was determined in 124 surgically resected NSCLC cases using methylation-specific polymerase chain reaction. RESULTS: The methylation frequencies of the genes tested in NSCLC specimens were 52% for E cadherin (CDH1), 41% for RAS association domain family protein (RASSF1A), 38% for fragile histidine triad (FHIT) and adenomatous polyposis coli (APC), 27% for retinoic acid receptor beta (RARbeta) and H-cadherin (CDH13), 20% for p16INK4A, 0.8% for O6-methylguanine-DNA-methyltransferase (MGMT), and 0% for glutathione S transferase P1 (GSTP1). The survival of the patients with FHIT methylation positive tumors was found to be significantly shorter than that for those patients with methylation-negative tumors (P=0.03), even in those patients with International Union Against Cancer TNM Stage I or Stage II disease (P=0.007). In contrast, there were no significant survival differences noted between the methylation-positive and methylation-negative tumors for the other genes tested. In addition, based on multivariate analyses, FHIT methylation-positive status was found to be independently associated with poor survival (P=0.046) and disease stage (P<0.0001). CONCLUSIONS: The results of the current study suggest that methylation of FHIT is a useful biomarker of biologically aggressive disease in patients with NSCLC. PMID- 15042682 TI - Biochemotherapy in patients with metastatic anorectal mucosal melanoma. AB - BACKGROUND: Patients with metastatic anorectal melanoma generally have an unfavorable prognosis, but no effective systemic therapy has been reported. METHODS: The authors retrospectively evaluated the medical records of all patients with metastatic anorectal melanoma treated with biochemotherapy between January 1991 and December 2001 at the University of Texas M. D. Anderson Cancer Center (Houston, TX). RESULTS: The search yielded 18 patients. Of these patients, 14 had undergone treatment with cisplatin (CDDP), vinblastine (VB), dacarbazine (DTIC), interferon alpha-2b (IFN), and interleukin 2 (IL-2); 2 had undergone treatment with CDDP, VB, DTIC, and IFN; 1 had undergone treatment with CDDP, IFN, and IL-2; and 1 had undergone treatment with CDDP, VB, temozolomide, IFN, and IL 2. All IL-2 treatments were administered intravenously. The median follow-up time was 12.2 months (range, 3.5-43.7 months). Eight patients (44%) had major responses, including two (11%) complete responses (CRs). Three patients were lost to follow-up evaluation after the completion of treatment. The median time to progression among the 15 remaining patients was 6.2 months. Four patients, including 1 with a CR, were alive at their last documented follow-up visits (survival: 14.0, 20.7, 31.3, and 43.7 months, respectively). The median overall survival was 12.2 months. Among 13 patients who received biochemotherapy as first line systemic therapy, 6 patients (46%) had major responses, including two (15%) CRs. The median time to progression for this group was 6.2 months, and the median overall survival was 12.9 months. CONCLUSIONS: Biochemotherapy had substantial activity against metastatic anorectal melanoma and should be considered for use in the treatment of metastatic disease from primary anorectal melanoma. PMID- 15042683 TI - 5-fluorouracil-induced oligodendrocyte death and inhibitory effect of cycloheximide, Trolox, and Z-VAD-FMK in murine cortical culture. AB - BACKGROUND: 5-fluorouracil (5-FU) is a widely used anticancer drug. One of the adverse effects of this drug is selective cerebral white matter injury, but to the authors' knowledge its mechanism has not been well documented. The current study was performed to investigate the mechanism of cerebral white matter injury caused by 5-FU and to develop the intervention to attenuate its injury in vitro. METHODS: Mixed oligodendrocyte/astrocyte cells were dissociated from specimens taken from approximately 2-day-old postnatal mouse cortex and cultured for 3-4 weeks. The culture cells were exposed to 5-FU, cycloheximide, emetine, Z-VAD-fmk, 2,3-dihydroxy-6-nitro-7-sulfamoyl-benzo(F)-quinoxaline (NBQX), Trolox, and epigallocatechin gallate. Oligodendrocyte cell death was assessed by counting the number of viable galactocerebroside-positive cells per x 100 field. RESULTS: Mixed oligodendrocyte/astrocyte culture cells that were exposed to 5-FU (at doses of 10 microM, 30 microM, and 100 microM) for 24 hours ensued concentration dependent oligodendrocyte death. The majority of oligodendrocytes, but few astrocytes, were injured by 100 microM 5-FU. Trolox, a vitamin E analog antioxidant, as well as cycloheximide (a protein synthesis inhibitor) and Z-VAD fmk (a caspase inhibitor), significantly attenuated the 5-FU-induced oligodendrocyte death. However, NBQX, an alpha-amino-2,3-dihydro-5-methyl-3-oxo-4 isoxazolepropionic acid (AMPA) receptor antagonist, did not appear to effect the 5-FU-induced oligodendrocyte death. CONCLUSIONS: The findings of the current study suggested that 5-FU led to oligodendrocyte death rather than astrocyte death by way of the apoptotic process, whereas antioxidants may prevent the 5-FU induced oligodendrocyte cell death in vitro. PMID- 15042684 TI - Intracranial meningeal hemangiopericytoma: the role of radiotherapy: report of 29 cases and review of the literature. AB - BACKGROUND: The current retrospective study was undertaken to evaluate the treatment outcomes of patients with meningeal hemangiopericytoma (M-HPC), to define the role of radiotherapy in the management of the disease, and to report on the patterns of failure. METHODS: The medical records of 29 patients with nonmetastatic M-HPC treated at The University of Texas M. D. Anderson Cancer Center (Houston, TX) between August 1979 and March 1999 were reviewed. Fifteen patients (52%) underwent macroscopic total resection ('gross total resection' [GTR]), 10 (34%) underwent subtotal resection (STR), and 4 (14%) had unknown extent of surgery. Ten of 24 patients received adjuvant radiotherapy. RESULTS: The 5, 10, and 15-year overall survival rates were 85%, 68%, and 43%, respectively. The local control rates at 5 and 10 years were 68% and 22%, respectively. Two patients experienced disease recurrence at another intracranial site. Sixteen patients (55%) developed extraneural metastases. Four patients (14%) developed spinal metastases. The 5-, 10-, and 15-year distant metastasis free survival rates were 80%, 46%, and 21%, respectively. The 5-year local control rates for patients treated with GTR and STR were 84% and 38%, respectively (P=0.003). Of the 15 patients treated with GTR, 3 received adjuvant radiotherapy as part of their initial treatment. Malignant disease did not recur locally in these three patients. However, the effect of the adjuvant radiotherapy on local control was not statistically significant. CONCLUSIONS: M-HPCs can recur locally or distantly in the neural axis or as extraneural distant metastases. Based on literature review and the patterns of failure in the current series, attempting to perform GTR followed by limited-field radiotherapy appeared to represent a reasonable approach for the initial management of M-HPC. PMID- 15042685 TI - Sequential dose-dense doxorubicin and ifosfamide for advanced soft tissue sarcomas: a Phase II trial by the Spanish Group for Research on Sarcomas (GEIS). AB - BACKGROUND: Combinations of high-dose ifosfamide (IF; 10-12 g/m2) plus doxorubicin (DX; 50-90 mg/m2) have been administered to patients with advanced soft tissue sarcoma (ASTS) in an attempt to improve therapeutic efficacy. Although these combination regimens appear to yield higher response rates than do standard-dose regimens, they also are associated with significant hematologic toxicity, despite the administration of hematopoietic growth factor support. As a potentially less toxic alternative, the authors designed a sequential, dose-dense schedule of DX and IF and explored its feasibility and toxicity, as well as patient compliance with the schedule, in a Phase II trial. METHODS: Chemotherapy naive patients with unresectable locally advanced or metastatic ASTS were to receive DX at 30 mg/m2 per day for 3 consecutive days once every 2 weeks for 3 cycles followed by IF at 12.5 g/m2 delivered by continuous infusion over 5 days once every 3 weeks for 3 cycles. Granulocyte-colony-stimulating factor was administered subcutaneously for 7 days beginning 24 hours after the completion of each DX or IF cycle. Additional IF cycles were allowed if an objective response was achieved. RESULTS: Sixty patients were enrolled in the trial. Three were ineligible, 9 had locally advanced disease, and 48 had metastatic disease. At the completion of therapy, the mean dose intensities for DX and IF were 40 mg/m2 per week and 3.87 g/m2 per week, respectively. Sixty-six percent of patients completed the regimen projected by the protocol. Grade 3 or 4 granulocytopenia, febrile neutropenia, and stomatitis occurred in 46%, 24%, and 27% of patients, respectively. Twenty of 53 assessable patients (38%; 95% confidence interval [CI], 25-51%) achieved objective responses, with a median time to progression of 24 weeks (95% CI, 18-30 weeks). CONCLUSIONS: Sequential administration of dose dense DX and high-dose IF is feasible and exhibits an acceptable hematologic toxicity profile and a level of activity that is within the range described for schedules that combine high-dose IF with an anthracycline. PMID- 15042686 TI - Familial association of specific histologic types of ovarian malignancy with other malignancies. AB - BACKGROUND: Population-based data on the familial association of specific histologic types of ovarian malignancy with other malignancies are limited. Such data may help to elucidate etiologic differences among histologic types of ovarian malignancy. METHODS: The nationwide Swedish Family-Cancer Database, which includes 10.3 million individuals and 20,974 ovarian carcinomas, was used to calculate standardized incidence ratios and 95% confidence intervals for age- and histology-specific ovarian malignancies in women whose parents or siblings were affected with malignancies at the most common disease sites. RESULTS: Ovarian malignancy was found to be associated with ovarian, laryngeal, breast, endometrial, liver, and colon carcinoma, as well as myeloma; epithelial ovarian malignancy was found to be associated with ovarian, endometrial, and skin malignancies and with melanoma and myeloma; papillary serous cystadenocarcinoma was found to be associated with ovarian and skin malignancies and with myeloma; and endometrioid carcinoma was found to be associated with endometrial, ovarian, and prostate malignancies and with melanoma. For younger women (ages 40-45 years) whose mothers were affected with endometrial malignancies, the risk of developing endometrioid carcinoma was slightly greater than the risk of developing papillary serous cystadenocarcinoma. CONCLUSIONS: Specific types of ovarian malignancy may be associated with specific familial disease sites, with such associations depending on age at diagnosis; the strength of the observed associations varied according to histology. Associations were found between endometrioid carcinoma and endometrial malignancy and between serous carcinoma and Hodgkin disease. PMID- 15042687 TI - Anthropometric variables, physical activity, and incidence of ovarian cancer: The Iowa Women's Health Study. AB - BACKGROUND: Reports on the relation between anthropometric variables (height, weight) and physical activity with ovarian cancer have been inconclusive. The objective of the current study was to extend investigation of potential associations in the Iowa Women's Health Study cohort. METHODS: The relation between self-reported anthropometric variables and incident ovarian cancer was studied in a prospective cohort of women ages 55-69 years who were followed for 15 years. Two hundred twenty-three incident cases of epithelial ovarian cancer were identified by linkage to a cancer registry. RESULTS: No association was found overall between ovarian cancer and height, but a positive association was observed for serous ovarian cancers (relative risk [RR], 1.86 for highest quartile vs. lowest quartile; 95% confidence interval [95% CI], 1.06-3.29). Although current body mass index (BMI) was not associated with ovarian cancer, a BMI > or =30 kg/m2 at age 18 years appeared to be associated positively with ovarian cancer (multivariate-adjusted RR, 1.83 for BMI > or =30 kg/m2 vs. BMI <25 kg/m2; 95% CI, 0.90-3.72), and this association was stronger after exclusion of the first 2 years of follow-up (RR, 2.15; 95% CI, 1.05-4.40). In a multivariate analysis, waist-to-hip ratio was associated with ovarian cancer (RR, 1.59 for highest quartile vs. lowest quartile; 95% CI, 1.05-2.40), but a linear dose response was not found. An index that combined the frequency and intensity of leisure-time physical activity was associated positively with ovarian cancer incidence (multivariate-adjusted RR, 1.42 for high activity vs. low activity; 95% CI, 1.03-1.97). This association was particularly strong for frequency of vigorous physical activity (multivariate-adjusted RR, 2.38 for >4 times per week vs. rarely/never; 95% CI, 1.29-4.38). CONCLUSIONS: Anthropometric variables were not major risk factors for ovarian cancer in the cohort studied; however, high BMI in early adulthood and frequent and vigorous physical activity may increase the risk of ovarian cancer among postmenopausal women. PMID- 15042688 TI - The use of complementary and alternative medical providers by insured cancer patients in Washington State. AB - BACKGROUND: Insurance coverage of complementary and alternative medicine (CAM) is expanding. However, to the authors' knowledge, little is know concerning CAM utilization among cancer patients under the insurance model of financing. In this study, the authors evaluated CAM provider utilization by cancer patients in a state that requires the inclusion of alternative practitioners in private, commercial insurance products. METHODS: An analysis was carried out of year 2000 claims data from two large Washington State insurance companies. RESULTS: Of 357,709 claimants, 7915 claimants (2.3%) had a cancer diagnosis. Among cancer patients, 7.1% had a claim for naturopathy, acupuncture, or massage; and 11.6% had a claim for chiropractic during the study year. The use of naturopathy (odds ratio [OR], 2.0; P<0.001) and acupuncture (OR, 1.4; P<0.001) were more common, and the use of chiropractic was less common (OR, 0.9; P<0.001) for cancer patients compared with those without cancer. No significant differences were noted in the use of massage between the two groups. Except in 2 individuals (0.03%), cancer patients also had at least 1 conventional provider claim during the year. Factors associated with nonchiropractic alternative provider use were female gender, the presence of metastatic cancer, hematologic malignancy, and the use of chemotherapy. Increased use of naturopathic physicians accounted for much of this trend. Musculoskeletal pain was the most common diagnosis at the CAM provider visit. Billed amounts for alternative services were <2% of the overall medical bills for cancer patients. CONCLUSIONS: A substantial number of insured cancer patients will use alternative providers if they are given the choice. The cost of this treatment is modest compared with conventional care charges. For individuals with cancer, CAM providers do not appear to be replacing conventional providers but instead are integrated into overall care. PMID- 15042689 TI - Strongyloidiasis in patients at a comprehensive cancer center in the United States. AB - BACKGROUND: The frequency of Strongyloides stercoralis infestation and complication in patients with cancer in the United States is unknown. METHODS: The authors performed a retrospective analysis of S. stercoralis infection in patients who were undergoing cancer treatment at The University of Texas M. D. Anderson Cancer Center (Houston, TX). RESULTS: The overall S. stercoralis infection frequency was approximately 1.0 per 10,000 new cancer cases between 1971 and 2003. Twenty-two of 25 patients (88%) were U.S. residents (19 from Texas; 1 each from Mississippi, Tennessee, and Puerto Rico), and the remaining 3 (13%) were from Latin America. Thirteen (52%) had solid-organ malignancies, whereas 12 (48%) had hematologic malignancies (lymphoma or multiple myeloma, n=8; leukemia, n=3; aplastic anemia, n=1). Twelve patients (48%) received systemic corticosteroids, 9 (36%) received antineoplastic therapy, and 2 underwent hematopoietic stem cell transplantation (HSCT). Diarrhea was reported in 13 patients (57%), and eosinophilia was observed in 11 patients (48%); 4 patients (16%) had probable hyperinfection syndrome (in 3 cases of polymicrobial gram negative bacteremia, 1 patient had Klebsiella pneumoniae pneumonia, whereas 1 patient presented with K. pneumoniae lung infection alone). Evidence of definite pulmonary hyperinfection syndrome was observed in 2 HSCT recipients (8%). Fourteen (74%) of 19 patients responded to thiabendazole therapy. Two patients with definite pulmonary hyperinfection syndrome developed fatal S. stercoralis hemorrhagic alveolitis despite receiving high-dose thiabendazole plus ivermectin therapy. CONCLUSIONS: In the current study, strongyloidiasis was uncommon in patients with cancer and remained localized in individuals with solid-organ malignancies. Definite pulmonary accelerated autoinfections were observed only in HSCT recipients. Therefore, pre-HSCT S. stercoralis screening in individuals from endemic regions of the United States warrants further study. PMID- 15042690 TI - Risk perception and concern among brothers of men with prostate carcinoma. AB - BACKGROUND: It is important for clinicians, researchers, and others who shape public health policy to understand the demographic correlates and psychologic factors that drive health behaviors, such as screening for early detection of cancer, particularly among individuals at high risk for developing the disease. METHODS: One-hundred eleven men whose brothers were diagnosed with prostate carcinoma completed a computer-assisted telephone interview aimed to assess their perception of absolute risk and concern about developing prostate carcinoma over the next 10 years and across their lifetime. Comparisons were made between selected demographic, behavioral, family pedigree characteristics, and measures of perceived risk and concern. RESULTS: The majority of men perceived their personal risk of developing prostate carcinoma to be > or =50%. Men who at the time of the interview were younger than their affected brother were significantly more concerned about prostate carcinoma and perceived their risk to be higher than men who were older than their brother. Estimates of personal risk and concern were also uniformly higher among men with more than one first-degree relative affected with prostate carcinoma compared to men with only one affected first-degree relative. Risk perception and concern about an impending prostate carcinoma diagnosis were associated with the use of supplements marketed for prostate health. CONCLUSIONS: The findings indicated that birth order in relation to a brother diagnosed with prostate carcinoma is significantly associated with risk perception and concern in unaffected family members. These results highlight the need for further study of the familial dynamics and characteristics that drive health behaviors and stress importance of public health education to inform men of personal risk assessment as well as the risks and benefits of screening. These studies ultimately can contribute to the success of strategies for the primary prevention and early detection of cancer. PMID- 15042691 TI - A randomized, active-control, pilot trial of front-loaded dosing regimens of darbepoetin-alfa for the treatment of patients with anemia during chemotherapy for malignant disease. PMID- 15042693 TI - Outpatient, sequential, parenteral-oral antibiotic therapy for lower risk febrile neutropenia in children with malignant disease: a single-center, randomized, controlled trial in Argentina. PMID- 15042695 TI - PI3K inhibitors block skeletogenesis but not patterning in sea urchin embryos. AB - Skeletogenesis in the sea urchin embryo is a simple model of biomineralization, pattern formation, and cell-cell communication during embryonic development. The calcium carbonate skeletal spicules are secreted by primary mesenchyme cells (PMCs), but the skeletal pattern is dictated by the embryonic ectoderm. Although the process of skeletogenesis is well characterized, there is little molecular understanding of the basis of patterning within this system. In this study, we examined the contribution of phosphatidylinositide 3-kinase (PI3K)-mediated signaling to the skeletogenic process in sea urchin embryos by using the well established PI3K inhibitors LY294002 and wortmannin. Our results show that PI3K inhibitors specifically and reversibly block skeletogenesis, and that this blockade occurs within the PMCs rather than in the ectoderm, because the inhibitors block spiculogenesis in cultured micromeres. Our results are consistent with a model in which PI3K signaling is required, not for pattern sensing or interpretation but rather for the biomineralization process itself in the sea urchin embryo. PMID- 15042696 TI - Sonic hedgehog signaling plays an essential role during embryonic salivary gland epithelial branching morphogenesis. AB - Gene targeting studies indicate that sonic hedgehog (Shh) signaling plays an essential role during craniofacial development. Because numerous mandibular derivatives (e.g., teeth, tongue, Meckel's cartilage) are absent in Shh null mice and the embryonic submandibular salivary gland (SMG) develops from the mandibular arch, we postulated that Shh signaling is important for embryonic SMG development. To address this question, we first determined the spatiotemporal distribution of Shh; two transmembrane proteins, patched 1 (Ptc) and Smoothened (Smo), which act as a negative or a positive regulator of the Shh signal, respectively; and the Gli 3 transcription factor, which is downstream of the Shh signal. The epithelial localization of Shh, Ptc, Smo, and Gli 3 suggests that Shh signaling may act within the epithelium in a juxtacrine manner. The SMG phenotype in our embryonic day (E) 18.5 Shh null mice can be characterized as "paedomorphic," that is, it fails to progress to ontogenic stages beyond the Early Pseudoglandular ( approximately E14). In a complementary set of experiments, we used organ culture to evaluate the effect of enhanced or abrogated Shh signaling on embryonic SMG development in vitro. Paired E13 (Late Initial Bud stage) or E14 (Pseudoglandular stage) SMGs were cultured in the presence or absence of exogenous Shh peptide supplementation; Shh-supplemented explants exhibit a significant stage-dependent increase in branching morphogenesis compared with control explants. Furthermore, by using cyclopamine, a steroidal alkaloid that specifically disrupts the Shh pathway, to abrogate endogenous Shh signaling in vitro, we found a significant decrease in branching in cyclopamine-treated explants compared with controls, as well as a significant decrease in epithelial cell proliferation. Our results indicate that Shh signaling plays an essential role during embryonic SMG branching morphogenesis. Exogenous FGF8 peptide supplementation in vitro rescues the abnormal SMG phenotype seen in cyclopamine-treated explants, demonstrating that overexpression of a parallel, but related, downstream signaling pathway can compensate for diminished Shh signaling and restore embryonic SMG branching morphogenesis. PMID- 15042697 TI - Identification and characterisation of mRif1: a mouse telomere-associated protein highly expressed in germ cells and embryo-derived pluripotent stem cells. AB - We have identified a mouse ortholog of the yeast Rif1 family of telomere associated proteins on the basis of its high expression in primordial germ cells and embryo-derived pluripotent stem cell lines. mRif1 is also highly expressed in totipotent and pluripotent cells during early mouse development, and in male and female germ cells in adult mice. mRif1 expression is induced during derivation of embryonic stem cells and is rapidly down-regulated upon differentiation of embryonic stem cells in vitro. Furthermore, we show that mRif1 physically interacts with the telomere-associated protein mTrf2 and can be cross-linked to telomeric repeat DNA in mouse embryonic stem cells. mRif1 may be involved in the maintenance of telomere length or pluripotency in the germline and during early mouse development. PMID- 15042698 TI - Myofibrillogenesis in the developing chicken heart: role of actin isoforms and of the pointed end actin capping protein tropomodulin during thin filament assembly. AB - Recently, important differences between myofibrillogenesis in cultured cardiomyocytes vs. the three-dimensional setting in situ could be determined. We investigated thin filament assembly in situ by confocal microscopy of whole-mount preparations of immunostained embryonic chicken hearts. Of interest, a distinct localisation of different actin isoforms was observed in immature thin filaments. Cardiac alpha-actin is restricted to filaments with a length comparable to mature thin filaments as soon as the first contractions occur, while vascular alpha actin makes up filaments that extend toward the M-band. The pointed-end actin filament capping protein tropomodulin can be found initially in close association with the plasma membrane, but attains its mature localisation pattern at the ends of the thin filaments only comparatively late during myofibrillogenesis. Thus tropomodulin acts as a length stabilising element of actin filaments also in developing cardiomyocytes in situ, but plays an additional role together with membrane-associated actin filaments in the earliest steps of myofibril assembly. PMID- 15042699 TI - Notch signaling can regulate endoderm formation in zebrafish. AB - Early in vertebrate development, the processes of gastrulation lead to the formation of the three germ layers: ectoderm, mesoderm, and endoderm. The mechanisms leading to the segregation of the endoderm and mesoderm are not well understood. In mid-blastula stage zebrafish embryos, single marginal cells can give rise to both endoderm and mesoderm (reviewed by Warga and Stainier [2002] The guts of endoderm formation. In: Solnica-Krezel L, editor. Pattern formation in zebrafish. Berlin: Springer-Verlag. p 28-47). By the late blastula stage, however, single marginal cells generally give rise to either endoderm or mesoderm. To investigate this segregation of the blastoderm into cells with either endodermal or mesodermal fates, we analyzed the role of Notch signaling in this process. We show that deltaC, deltaD, and notch1 are expressed in the marginal domain of blastula stage embryos and that this expression is dependent on Nodal signaling. Activation of Notch signaling from an early stage leads to a reduction of endodermal cells, as assessed by sox17 and foxA2 expression. We further find that this reduction in endoderm formation by the activation of Notch signaling is preceded by a reduction in the expression of bonnie and clyde (bon) and faust/gata5, two genes necessary for endoderm formation (Reiter et al. [1999] Genes Dev 13:2983-2995; Reiter et al. [2001] Development 128:125-135; Kikuchi et al. [2001] Genes Dev 14:1279-1289). However, activation of Notch signaling in bon mutant embryos leads to a further reduction in endodermal cells, also arguing for a bon-independent role for Notch signaling in endoderm formation. Altogether, these results suggest that Notch signaling plays a role in the formation of the endoderm, possibly in its segregation from the mesoderm. PMID- 15042700 TI - T-box transcription factor Tbx2 represses differentiation and formation of the cardiac chambers. AB - Specific regions of the embryonic heart tube differentiate into atrial and ventricular chamber myocardium, whereas the inflow tract, atrioventricular canal, inner curvatures, and outflow tract do not. These regions express Tbx2, a transcriptional repressor. Here, we tested its role in chamber formation. The temporal and spatial pattern of Tbx2 mRNA and protein expression in mouse hearts was found to be complementary to that of chamber myocardium-specific genes Nppa, Cx40, Cx43, and Chisel, and was conserved in human. In vitro, Tbx2 repressed the activity of regulatory fragments of Cx40, Cx43, and Nppa. Hearts of transgenic embryos that expressed Tbx2 in the prechamber myocardium completely failed to form chambers and to express the chamber myocardium-specific genes Nppa, Cx40, and Chisel, whereas other cardiac genes were normally expressed. These findings provide the first evidence that Tbx2 is a determinant in the local repression of chamber-specific gene expression and chamber differentiation. PMID- 15042701 TI - Atlas of Hox gene expression in the developing kidney. AB - Hox genes often play important roles in segment identity determination and organogenesis. To better understand the roles of Hox genes during kidney development, we performed an extensive analysis of their expression patterns. Section in situ hybridizations were used to define the expression of 37 Hox genes at embryonic day (E) 12.5, E13.5, E15.5, and E17.5 of kidney development. Several interesting principles emerged. First, the concept of colinearity was preserved. Hox genes from the more 3' positions in clusters were more often expressed in the ureteric bud, which is derived from the anterior of the intermediate mesoderm. Second, Hox genes were expressed throughout the ureteric bud without any segment specificity. Third, in the different segments of the forming nephron we did observe overlapping domains of Hox gene expression, which initiated distally at the junction between the nephron and ureteric bud, and extended proximally variable distances. Finally, we observed that paralogous Hox genes often showed surprisingly diverse expression patterns. Indeed, contiguous genes on a single cluster more often showed similar expression patterns than paralogs. In summary, the resulting atlas of Hox gene expression provides a foundation for further study of the overlapping functions Hox genes in the developing kidney. PMID- 15042702 TI - Functional analysis in Drosophila indicates that the NBCCS/PTCH1 mutation G509V results in activation of smoothened through a dominant-negative mechanism. AB - Mutations in the human homolog of the patched gene are associated with the developmental (and cancer predisposition) condition Nevoid Basal Cell Carcinoma Syndrome (NBCCS), as well as with sporadic basal cell carcinomas. Most mutations that have been identified in the germline of NBCCS patients are truncating or frameshift mutations, with amino acid substitutions rarely found. We show that a missense mutation in the sterol-sensing domain G509V acts as a dominant negative when assayed in vivo in Drosophila. Ectopic expression of a Drosophila patched transgene, carrying the analogous mutation to G509V, causes ectopic activation of Hedgehog target genes and ectopic membrane stabilisation of Smoothened. The G509V transgene behaves in a manner similar, except in its subcellular distribution, to a C-terminal truncation that has been characterised previously as a dominant negative protein. G509V exhibits vesicular localisation identical to the wild type protein, but the C-terminal truncated Patched molecule is localised predominantly to the plasma membrane. This finding suggests that dominant negative function can be conferred by interruption of different aspects of Patched protein behaviour. Another mutation at the same residue, G509R, did not exhibit dominant-negative activity, suggesting that simple removal of the glycine at 509 is not sufficient to impart dominant-negative function. PMID- 15042703 TI - Protein phosphatase 1alpha is required for murine lung growth and morphogenesis. AB - Protein phosphatase 1 (PP1) plays important roles in cell cycle control and apoptosis, two processes that impinge on morphogenesis and differentiation. Following the precedent set by other molecules regulating the cell cycle and apoptosis, we hypothesized that PP1 may have context-specific roles in development. Therefore, we have studied the spatial and temporal expression of PP1alpha during murine lung development and determined the consequences of loss of PP1alpha function on branching morphogenesis. By using an immunohistochemical approach, we show here that PP1alpha was expressed throughout the epithelium and mesenchyme upon the emergence of the lung primordium on embryonic day 10, with immunostaining exclusively extranuclear. During the late pseudoglandular stage, PP1alpha was predominantly expressed in the distal lung epithelium, whereas the mesenchyme contained very little or no PP1alpha protein. Peri- and postnatally, PP1alpha immunostaining was mostly nuclear in apparently differentiated cells, as judged by colocalization with well-known markers for lung differentiation. Exposure of fetal lung explants to antisense oligodeoxynucleotides against PP1alpha, resulted in decreased overall size of the cultured lung, a defect in forming new airways, lack of expression of surfactant protein C, and histologic signs of poor differentiation. These data suggest that PP1alpha is required for branching morphogenesis and differentiation. PMID- 15042704 TI - Cellular growth and rearrangement during the development of the mammalian organ of Corti. AB - The sensory epithelium of the mammalian cochlea, the organ of Corti, is comprised of ordered rows of cells, including inner and outer hair cells. Recent results suggest that physical changes in the overall size and shape of the cochlear duct, including possible convergence and extension, could play a role in the development of this pattern. To examine this hypothesis, changes in cell size and distribution were determined for different regions of the cochlea duct during embryonic development. In addition, changes in the spatial distribution of sensory precursor cells were determined at different developmental time points based on expression of p27kip1. Unique changes in luminal surface area, cell density, and number of cell contacts were observed for each region of the duct. Moreover, the spatial distribution of p27kip1-positive cells changed from short and broad early in development, to long and narrow. These results are consistent with the hypothesis that convergence and extension plays a role in cellular patterning within the organ of Corti. PMID- 15042705 TI - Characterization of c-Kit and nestin expression during islet cell development in the prenatal and postnatal rat pancreas. AB - It has been well documented that there are abundant endocrine progenitor cells in the neonatal pancreas. However, little is known of their relative proportions or even their phenotypes. The aim of this study was to examine the normal distribution and characteristics of putative endocrine precursor cells, identified by c-Kit or nestin expression, within the prenatal and postnatal rat pancreas during islet cell development. Here, we provide evidence of the existence of a subset of ductal, islet, and acinar cells with an immature morphology and high proliferative capacity that expressed c-Kit or nestin. The proportion of islet cells expressing c-Kit or nestin was highest at embryonic day 18 (25 +/- 4% and 28 +/- 6%) and decreased significantly by postnatal day 28 (P < 0.01), 1.3 +/- 0.2% and 5.7 +/- 1%, respectively. The expression of nestin mRNA decreased throughout development, while c-Kit mRNA expression was found to slightly increase in the developing pancreas. Coexpression patterns indicated that c-Kit and nestin form two distinct cell populations in the postnatal pancreas, and infrequently coexpress with other pancreatic cell-specific markers. Furthermore, decreased c-Kit and nestin expression in the islets in postnatal life correlated with an increase in cells immunopositive for Pdx-1 compared with birth (36 +/- 5% vs. 60 +/- 3%, P < 0.01), which accompanied a doubling in the proportion of Glut-2-positive cells (39.4 +/- 4% vs. 68.8 +/- 3%, P < 0.01), both of which are mature beta-cell markers. Taken together, these findings suggest that c-Kit- and nestin-expressing cells represent endocrine precursor cells that undergo marked changes in population dynamics during the transition from prenatal to postnatal pancreatic development in the rat. Characterization of the phenotype, relative abundance and location of these cells within the developing pancreas is an important step toward creating a strategy for isolating stem cell populations and modeling islet cell differentiation in vitro. PMID- 15042706 TI - Knock-in of nuclear localised beta-galactosidase reveals that the tyrosine phosphatase Ptprv is specifically expressed in cells of the bone collar. AB - Ptprv is a member of the transmembrane tyrosine phosphatase gene family reported to be expressed in osteoblasts and gonads. To better define the developmental and tissue specificity of Ptprv expression, we generated knock-in mice expressing a nuclear localised beta-galactosidase reporter under the control of resident Ptprv regulatory elements. Histochemical staining of Ptprv-nLacZ mice revealed that Ptprv expression is readily detectable in the foetal gonadal ridge of both sexes and in adult gonads where it is localised to Sertoli cells of the testis and celomic epithelial cells of the ovaries. During early limb development, Ptprv expression is prominent in the apical ectodermal ridge of the limb bud. At latter stages of development, Ptprv is predominantly expressed in the perichondrial and periosteal region of long bones, known as the bone collar. In contrast to previous indications from in vitro studies, there is little if any expression in mature osteoblasts in vivo. Analysis of Ptprv mRNA localisation by in situ hybridization in parallel with molecular markers of chondrocytes and osteoblasts confirmed the specific expression of Ptprv in immature bone collar cells. The specificity of Ptprv expression in these cells may be a useful tool to elucidate their role in the transition of skeletal elements from cartilage template to bone. PMID- 15042707 TI - nlz gene family is required for hindbrain patterning in the zebrafish. AB - This study describes the conserved nlz gene family whose members encode unusual zinc finger proteins. In the zebrafish neurectoderm, both nlz1 and the newly isolated nlz2 are expressed in the presumptive hindbrain and midbrain/hindbrain boundary, where expression of nlz1 is dependent on pax2a. In addition, nlz2 is uniquely expressed more anteriorly, in the presumptive midbrain and diencephalon. Overexpression of Nlz proteins during gastrula stages inhibits hindbrain development. In particular, ectopically expressed Nlz1 inhibits formation of future rhombomeres 2 and 3 (r2, r3), whereas neighboring r1 and r4 are not affected. Conversely, simultaneous reduction of Nlz1 and Nlz2 protein function by expression of antisense morpholino-modified oligomers leads to expansion of future r3 and r5, with associated loss of r4. These data indicate that one function of the nlz gene family is to specify or maintain r4 identity, and to limit r3 and r5 during hindbrain formation. PMID- 15042708 TI - Up-regulation of muscle-specific transcription factors during embryonic somitogenesis of zebrafish (Danio rerio) by knock-down of myostatin-1. AB - Myostatin, a secreted growth and differentiation factor (GDF-8) belongs to transforming growth factor (TGF-beta) superfamily that plays as a negative regulator of skeletal muscle development and growth. Recently, myostatin has been isolated from fish; however, its role in muscle development and growth remains unknown. Here, we present the expression of myostatin during development and the effects of its knock-down on various genes such as muscle regulatory transcription factors (MRFs), muscle-specific proteins (MSP), and insulin-like growth factors (IGFs). The myostatin expression was found to be maternal as it starts in one-cell stage onward. The reverse transcription-polymerase chain reaction (RT-PCR), in situ hybridization, and Southern and Northern blots demonstrated that the myostatin expression is not only restricted to skeletal muscle, but it expressed all the tested tissues. Expression of myostatin was effected by using antisense morpholinos resulted in significant phenotypic difference in stages 18 and 20 hours postfertilization (hpf). To confirm the specificity of myostatin morpholino, furthermore, a rescue experiment was conducted. The length as well as width of somites was increased with almost no gap in between the somites. In addition, it deserves to mention that this is a first animal model that shows changes in the size of the somites. Moreover, analyses of MRFs, MSP, and IGFs in the knock-down embryos by RT-PCR revealed the up-regulation of MyoD, Myogenin, and Mck transcription, whereas IGF-2 transcription showed mild response with no effect on IGF-1, Desmin, and Myf5. In situ hybridization showed that there was an increase in the number of somites from 3 to 4 at 13 and 22 hpf. Taken together, these data suggest that myostatin plays a major role during myogenesis, apart from inhibition of proliferation as well as differentiation. PMID- 15042709 TI - Periostin is expressed within the developing teeth at the sites of epithelial mesenchymal interaction. AB - Periostin was originally isolated as an osteoblast-specific factor that functions as a cell adhesion molecule for preosteoblasts and is thought to be involved in osteoblast recruitment, attachment, and spreading. The protein was renamed "periostin" because of its expression in the periosteum and periodontal ligament, indicating a potential role in bone and maintenance of tooth structure. Periostin has structural similarity to insect fasciclin-I and can be induced by TGF-beta and Bmp2. Because tooth and periodontium development is a well-described genetic model for organogenesis governed by a reciprocal set of epithelial-mesenchymal interactions, thought to be controlled by various TGF-beta superfamily members, we investigated whether periostin is present during tooth morphogenesis. Both periostin mRNA and protein expression were analyzed throughout normal tooth development (embryonic day [E] 9.5-newborn) and within both Bmp4- and Msx2-null embryos. Periostin mRNA is initially present within the E9.5 first branchial arch epithelium and then shifts to underlying ectomesenchyme. Both mRNA and protein are asymmetrically localized to the lingual/palatal and buccal side during the early epithelial-mesenchymal interactions. Periostin is also present in dental papilla cells and within the trans-differentiating odontoblasts during the bell and hard tissue formation stages of tooth development. We suggest that periostin plays multiple roles as a primary responder molecule during tooth development and may be linked to deposition and organization of other extracellular matrix adhesion molecules during maintenance of the adult tooth, particularly at the sites of hard-soft tissue interface. PMID- 15042710 TI - Developmental expression of the POU domain transcription factor Brn-3b (Pou4f2) in the lateral line and visual system of zebrafish. AB - Members of the class IV POU domain transcription factors are important regulators of neural development. In mouse, Brn-3b (Pou4f2, Brn3.2) and Brn-3c (Pou4f3, Brn3.1) are essential for the normal differentiation and maturation of retinal ganglion cells (RGCs) and hair cells of the auditory system, respectively. In this report, the cloning and expression profile of brn-3b in the zebrafish (Danio rerio) were assessed as the first step for understanding its role in the development of sensory systems. Two brn-3b alternative transcripts exhibited different onset of expression during development but shared overlapping expression domains in the adult visual system. The brn-3b expression in the zebrafish retina was consistent with a conserved role in differentiation and maintenance of RGCs. Expression was also observed in the optic tectum. Unexpectedly, brn-3b was prominently expressed in the migrating posterior lateral line primordium and larval neuromasts. For comparison, brn-3c expression was limited to the otic vesicle and was not detected in the lateral line during embryonic development. The expression of brn-3b in the mechanosensory lateral line of fish suggests a conserved function of a class IV POU domain transcription factor in sensory system development. PMID- 15042711 TI - Reciprocal chemokine receptor and ligand expression in the human placenta: implications for cytotrophoblast differentiation. AB - At the onset of pregnancy, the human placenta, which forms the interface between the embryo/fetus and the mother, must rapidly develop into a life-sustaining organ. The many unusual processes entailed in placental development include the poorly understood phenomenon of maternal tolerance of the hemiallogeneic cells of the conceptus, including, most remarkably, placental trophoblasts that invade the uterine wall. To investigate whether this fetal organ exerts control over the maternal immune system at the level of leukocyte trafficking, we examined placental expression of chemokines, well-known cytokine regulators of leukocyte movements. In situ hybridization revealed abundant expression of 13 chemokines in the stromal but not the trophoblast compartment of chorionic villi. Potential roles for these molecules include recruitment of the resident macrophage (Hofbauer cell) population to the villi. In parallel, cytotrophoblast production of a panel of nine chemokine receptors was assessed by using RNase protection assays. The numerous receptors detected suggested the novel possibility that the paracrine actions of chemokine ligands derived from either the villous stroma or the decidua could mediate general aspects of placental development, with specific contributions to cytotrophoblast differentiation along the pathway that leads to uterine invasion. PMID- 15042712 TI - Microarray analysis of retinoid-dependent gene activity during rat embryogenesis: increased collagen fibril production in a model of retinoid insufficiency. AB - Retinoic acid (RA) is an essential mediator of embryogenesis. Some, but not all, of its targets have been identified. We previously developed a rat model of gestational retinoid deficiency (RAD; Power et al. [1999] Dev. Dyn. 216:469-480) and generated embryos with developmental impairments that closely resemble genetic and dietary models of retinoid insufficiency. Here, we used microarray analysis and expression profiling to identify 88 transcripts whose abundance was altered under conditions of retinoid insufficiency, as compared with normal embryos. Among these, the induction by RAD of genes involved in collagen I synthesis (COL1A1, IA2 and VA2, prolyl-4-hydroxylase-alpha1) and protein galactosylation (galactokinase, ABO galactosyltransferase, UDP-galactose transporter-related protein) was especially noteworthy because extracellular matrix regulates many developmental events. We also identified several genes involved with stress responses (cathepsin H, UBC2E, IGFBP3, smoothelin). Real time polymerase chain reaction analysis of selected candidates revealed excellent agreement with the array findings. Further validation came from the demonstration that these genes were similarly dysregulated in two genetic models of retinoid insufficiency, the retinol binding protein null-mutant embryo and the Raldh2 null mutant embryo. In situ hybridization of RAD embryos found increased collagen IA1 and IGFBP3 mRNA within the connective mesenchyme and vasculature, respectively, and a failure to repress the growth factor midkine within the RAD neural tube. Many of the identified genes were not known previously to respond to retinoid status and will provide new insights to retinoid roles and to the consequences of retinoid insufficiency. PMID- 15042713 TI - Identification of a nonchordate-type classic cadherin in vertebrates: chicken Hz cadherin is expressed in horizontal cells of the neural retina and contains a nonchordate-specific domain complex. AB - Classic cadherins mediate calcium-dependent cell-cell adhesion in a variety of animals, but there are marked differences in their domain structures between chordate and nonchordate animals. The extracellular domain of chordate-type classic cadherins (type I and type II classic cadherins) consists of five tandem repeats of conserved sequences called EC domains, whereas that of nonchordate type classic cadherins (designated as type III classic cadherin) contains a variable number of EC domains, followed by a characteristic domain complex made of laminin-A globular domains and EGF-like repeats. In the present study, we identified a novel vertebrate type III cadherin showing high sequence similarity to Drosophila N-cadherin, and named this molecule chicken Hz-cadherin (cHz cadherin), because of the distinct expression in horizontal cells of the neural retina. cHz-cadherin functioned as an adhesion molecule when introduced into cultured cells. Database search revealed one cHz-cadherin homologue in zebrafish and two in puffer fish, but none in other vertebrate species examined. These observations indicate that type III classic cadherins have been conserved in vertebrate species, being expressed by limited cells types, but lost in particular phylogenic groups of the vertebrates. PMID- 15042715 TI - Proceedings of the 2nd International Meeting of Steroids and Nervous System. February 22-26, 2003, Turin, Italy. PMID- 15042714 TI - Genetic background has a major effect on the penetrance and severity of craniofacial defects in mice heterozygous for the gene encoding the nucleolar protein Treacle. AB - Treacher Collins syndrome (TCS) is a craniofacial disorder that results from mutations in TCOF1, which encodes the nucleolar protein Treacle. The severity of the clinical features exhibits wide variation and includes hypoplasia of the mandible and maxilla, abnormalities of the external ears and middle ear ossicles, and cleft palate. To determine the in vivo function of Treacle, we previously generated Tcof1 heterozygous mice on a mixed C57BL/6 and 129 background. These mice exhibited a lethal phenotype, which included abnormal development of the maxilla, absence of the eyes and nasal passages, and neural tube defects. Here, we show that placing the mutation onto different genetic backgrounds has a major effect on the penetrance and severity of the craniofacial and other defects. The offspring exhibit markedly variable strain-dependent phenotypes that range from extremely severe and lethal in a mixed CBA/Ca and 129 background, to apparently normal and viable in a mixed BALB/c and 129 background. In the former case, in addition to a profoundly severe craniofacial phenotype, CBA-derived heterozygous mice also exhibited delayed ossification of the long bones, rib fusions, and digit anomalies. The results of our studies indicate that factors in the different genetic backgrounds contribute extensively to the Tcof1 phenotype. PMID- 15042716 TI - Community pharmacists do not intentionally transmit incorrect claim information- but caution is warranted in days supply calculations. PMID- 15042717 TI - Community pharmacists do not intentionally transmit incorrect claim information- but caution is warranted in days supply calculations. PMID- 15042718 TI - The cutting edge. PMID- 15042719 TI - Listening to staff key for hospitals on Fortune's list of top employers. PMID- 15042720 TI - Patient-friendly, high-tech facilities a trend in OR construction projects. PMID- 15042721 TI - Change assistant pay, GAO advises. PMID- 15042722 TI - Penelope, the robot, could scrub in. PMID- 15042723 TI - Gaining efficiency with instrument tracking. PMID- 15042724 TI - Practical guide for improving performance. PMID- 15042725 TI - What's ASC's obligation for escorts? PMID- 15042726 TI - JCAHO tailors safety goals for ambulatory care. PMID- 15042727 TI - [Current problems in psychiatric health care in Poland (2003)]. AB - The paper is a presentation of the most important and actual psychiatric health care problems in Poland: the financial situation of hospitals, the risks faced by the out-patient psychiatric care, maintaining the legal issues of the psychiatric patients, the situation of forensic psychiatry and legal-psychiatric expertise, postgraduate training, availability of psychotropic drugs. Attention is paid to the importance of giving the programme of psychiatric health protection a status of evident value and that this programme should become a national programme. PMID- 15042728 TI - [Prevalence of depression in Krakow population of 17 years old students in years 1984 and 2001]. AB - Results of scarce epidemiological studies on depression in adolescents are diverse and dependent of survey methodology as well as on the theoretical approach of researchers. Those studies where the methods are comparable show that depression prevalence depends on the macrosocial situation and the adolescence stage. AIM: The study aimed to assess changes in depression prevalence in mid adolescents between 1984 and 2001. A presumption was made that the social situation in Poland had changed in the last 17 years. METHOD: In 2001 a representative sample of school attending 17 y.o. adolescents was screened with KID-IO"C1". The results were compared with those of a similar survey, using the same method, carried out in 1984. RESULTS: Point prevalence indexes appeared to be similar as well as higher for girls in comparison with boys. However, dependence of depression prevalence and the type of the senior school has changed. In 1984 it was higher in senior schools preparing for university studies, while in 2001--in those which train in a skill. CONCLUSIONS: Earlier conceptualisations of adolescent depression as connected with social conditions making ways to adulthood difficult (e.g. unemployment) as well as high indexes of depression prevalence in adolescents living in regions with higher unemployment rates than in Krakow, allow for an interpretation of the findings as result of macrosocial changes. Other hypotheses e.g. influence of the adolescence timing and course require a longitudinal prospective study. PMID- 15042729 TI - [Child maltreatment as a risk factor for suicidal behavior--a literature review]. AB - Based on a review of studies done in the last 20 years the author presents recent knowledge on the possible relationship between child maltreatment and suicidal behaviour in childhood, adolescence and adulthood. PMID- 15042730 TI - [Depressive disorders in adolescents with mild retardation. Research report]. AB - The clinicians agree to a point that mental retardation is a risk factor for the development of a mental disorder and that all kinds mental disorders are present in this population. AIM: The aim of this research was to find answers to two questions: 1. Was the intensity of depressiveness higher in adolescents with mild mind retardation in comparison to adolescents with normal IQ? 2. Were variables like gender, family situation and a religious attitude associated with the rate of depressiveness? METHOD: The subjects were 160 adolescents (80 with mild mind retardation and 80 with normal IQ). The Children's Depression Inventory by M. Kovacs was used. RESULTS: The results pointed out that an intensity of depressiveness was significantly higher in adolescents with mental retardation. In the whole population girls had a higher rate of depressiveness in comparison to boys and in the whole population the family situation influenced the results in CDI. In adolescents with mental retardation a religious attitude was not significantly associated with depressiveness. In adolescents with normal IQ this dependence was found. PMID- 15042731 TI - [Eating disorder prevention program--pilot study]. AB - AIM: Recent research has suggested that school-based programmes of prevention of eating disorders obtained ambiguous results. The aim of our work was to establish the efficacy of the eating disorders prevention programme for schoolgirls of secondary schools. METHOD: Subjects in the experimental groups received 8 sessions of practice with various tasks, while the control group did not participate in sessions of psychoeducation. Overall, we examined 109 adolescents participating in sessions and 117 adolescents in the control group. Participants were examined at the beginning of the programme, after the operation and 6 months later. Assessment was made with EDI, EAT-26. RESULTS: Assessment before and after the study phase showed that participants in the experimental groups had not made significantly more improvement than the control group. It was also shown that in the two subgroups (girls form technical college and girls form college) there were significant differences in the pre-test, post-test and follow-up results. CONCLUSIONS: Similarities that were obtained through the research suggest the necessity of examining other possible relationships between chosen variables. Analysis of participants' feedback information emphasises that the changes the programme brought appeared feasible in the social functioning area (with peers, parents relations). It is not excluded that it is too late for eating disorder prophylaxis in the phase of personal development, as some researchers argue. Differences in results between schoolgirls from college and from technical college suggest some differences in risk and protective factors. It seems important to verify this. PMID- 15042732 TI - [Prevention of eating disorders in children and adolescents. Part I. General considerations and risk factors]. AB - The paper presents general considerations concerned with using prevention programmes mental disorders in children and adolescents. In this context the authors try to describe current views on creating prevention programmes of eating disorders. These programmes require clearly defined, potentially modifiable risk factors and protective factors. PMID- 15042733 TI - [Prevention of eating disorders in children and adolescents. Part II. The effectiveness of prevention programs]. AB - This paper presents various models of child and adolescent prevention programmes of eating disorders with the emphasis on the efficiency of these models. On the ground of the available literature, the authors present the results of these methods. It seems that in every prevention programme risk and protective factors, which are connected with the given intervention should be precisely defined. Therefore it is absolutely necessary that the programmes should be clear with better inner coherence which will consequently result in better efficiency. PMID- 15042734 TI - [Evaluation of temperament and personality in bulimia nervosa]. AB - Previous studies revealed the possibility of abnormal personality development role in the etiology of eating disorders. It was found that a diagnosis of personality disorders, mostly borderline personality and/or histrionic personality can be made in about 44-46% of bulimic patients. The inconsistencies in identifying personality types using categorical assessment approaches have encouraged a conceptualization of the personality from a dimensional perspective. It was revealed that the Tridimensional Personality Questionnaire (TPQ) and Temperament and Character Inventory (TCI) are useful in studying patients with eating disorders. AIM: The aim of the study was personality dimensions and depression symptoms assessment in bulimic patients. METHOD: We studied 36 women with purging type bulimia according to DSM-IV and ICD 10 criteria. The mean age of the studied women was 19.7 years. The control group were 44 healthy women: university and last year high school students (mean age 20.4). We assessed body height, body mass and Body Mass Index (BMI). Severity of depression symptoms was assessed with the use of Beck Depression Index (BDI), and personality dimensions with the use of (TCI). CONCLUSIONS: Women from the study group had higher harm avoidance (HA) scores, which shows that bulimic patients are shy, fearful, doubtful, tend to be inhibited in most social situations. Mood disorders present in the studied women influenced HA scores. The results of the study revealed lower possibilities of self-directness in bulimic patients in comparison with the control group. Negative correlation between BDI and SD scores, shows that a decrease of depression severity may lead to an increase of self-esteem, independence and effectiveness of bulimic patients. PMID- 15042735 TI - [The role of orexines in appetite regulation]. AB - Anorexia nervosa (AN) belongs to the group of eating disorders. Many different factors are taken into consideration as far as the origin of this disorder is concerned, among them: individual factors (genetic, biological), personality factors, sociocultural factors, family factors. Among the biological factors, the role of neuropeptides is considered. Last few years (1998) have resulted in the discovery of two neuropeptides--orexines OXA and OXB which--apart from being regarded as appetite stimulants--are also supposed to be responsible for the energy metabolism of an organism. Orexines, a.k.a. hypocretins, arise from the disintegration of their common precursor--the polypeptide: preorexine. Their name derives from the Greek word for appetite: orexis. The orexine A (OXA) is a 33 amino acid peptide consisting of 2 chains connected by the Cys 6--Cys 12 and Cys 7--Cys 14 bridges. It is a potential food intake and gastric juice secretion stimulant. The connecting bridges in OXA play a crucial role in the receptor OXA 1 activation. The orexine B (OXB) is a 28 amino acid peptide with no connecting bridges between the chains. It mainly plays a role in the energy metabolism of an organism and does not influence the secretion of gastric juice. The OXR-1 receptor is a selective receptor for OXA, while the OXR-2 receptor is not privileged for any of the two orexines--both of them can connect to it. Orexines are produced by a small number of hypothalamus neurons, mainly in the lateral hypothalamus (LHA), but also the posterior hypothalamus--the so-called "eating center". Orexines control: the wakefulness-sleep state, food intake (OXA over 100 times stronger than OXB) and the neuroendocrine system. Their discovery may help in understanding the mechanism of anorexia nervosa. PMID- 15042736 TI - [Pediatric autoimmune neuropsychiatric disorders associated with streptococcal infections (PANDAS). A report of two cases]. AB - AIM: To critically review the past years of research on paediatric autoimmune neuropsychiatric disorders associated with streptococcal infections in children and adolescents. METHOD: Literature on PANDAS published from 1995 onward was systematically reviewed. The review focuses on definition and diagnostic consideration aetiological and therapeutical issues. To illustrate the clinical characteristic of PANDAS authors present two cases of children with a severe course of obsessive-compulsive disorder and Tourette's syndrome. RESULTS: Post streptococcal autoimmunity has been postulated as an aetiologic factor in the development of childhood-onset obsessive-compulsive disorder, tic disorders including Tourette's disorder. This hypothesis arose from a series of clinical observations including the documentation of obsessive-compulsive symptoms of children affected by Sydenham's chorea, a variant of rheumatic fever characterised by neurological dysfunction and also by concomitant investigations of childhood-onset OCD and Tourette's syndrome. CONCLUSION: Results of these studies led to the identification of children whose clinical course is characterised by abrupt and dramatic symptom exacerbations which are temporally related to group A beta-hemolytic streptococcal infections. The identification of such a subgroup will allow for testing of the model of pathogenesis as well as development of novel treatment and prevention strategies. Future research are needed to explore the nature of PANDAS and their relationship with different psychiatric disorders in children and adolescents. PMID- 15042737 TI - [Comprehensive treatment of nicotine dependence in patients with cardiovascular diseases--personal experience]. AB - AIM: To evaluate psychological factors influencing the effectiveness of smoking cessation using bupropion and psychological intervention in patients with cardiovascular diseases. METHOD: A group of 59 patients (26 M, 33 F; mean age 44.2 +/- 10.2 years) identified among smokers admitted to the 1st Cardiac Department between 2000 and 2001 with hypertension (n = 46) and coronary artery disease (n = 13). Entry criteria to participate in a 7-week programme included nicotine dependence and no contraindications to bupropion. Pharmacotherapy was used in parallel with psychological interventions based upon cognitive and behavioural methods. The type and degree of nicotine dependence was measured using Fragerstrom tolerance questionnaire whereas motivation was scored according to the Schneider questionnaire. The severity of depression symptoms was measured by the Beck Depression Inventory. RESULTS: At 2 months and at one year after discontinuation of treatment 74.6% and 40% of the patients respectively maintained abstinence from smoking. Among patients with good therapeutic effects the level of motivation to quit smoking was higher (p < 0.001 vs. p < 0.007). The level of motivation was correlated with the severity of depression symptoms (p < 0.005) but not with nicotine dependence, age and sex. Motivation to stop smoking is related to the depression symptoms (beta = -0.32; p = -.01). CONCLUSION: Patients with initially higher motivation benefit more from treatment of nicotine dependence. The level of motivation is adversely affected by the depressive mode of functioning (negative cognitive patterns, learned helplessness). It appears necessary to take into account the assessment of depressive disorders when planning treatment of nicotine dependence in patients with cardiovascular diseases. PMID- 15042738 TI - [Parasitic diseases in the Russian Federation under the conditions of population migration]. AB - Migration of the population has been established to substantially increase the risk of importation of parasitic diseases into Russia. Mass importation of malaria into Russia has started since 1993 when local cases of malaria (secondary to imported ones), whose source was newcomers from Tadjikistan and Azerbaijan, were notified. The migration of the population has increased the incidence of helminthiasis and protozoan diseases earlier sporadically recorded in Russia. In 1996 to 2002, a total of 7386 patients with parasitic diseases considered by official statistics as other helminthiasis and other protozoan diseases were notified. In some areas, the morbidity rates due to hymenolepidiasis and trichocephaliasis have been formed from imported cases, the number of imported cases of opisthorchiasis increased, the area of spreading echinococciasis in Russia expanded at the expense of imported cases of echinococciasis from Kazakhstan, Uzbekistan, and the Ukraine. PMID- 15042739 TI - [The parasitological situation in the Khanty-Mansi autonomous region]. PMID- 15042740 TI - [Imported cases of malaria and HIV infection in Russia]. AB - Malaria does not belong to a group of diseases that are markers of HIV infection; a combined transmission of malaria and HIV infection does not occur (except for cases of vertical and syringe-associated transmission of causative organisms), but HIV infection is more common in the malaria-endemic areas since both diseases are socially related. Expansion of both infections is associated with the same social processes in society. This paper analyzes the relationship of the spread of HIV-infection to the routes of migration of narcotic agents in case of Russia, by comparing the spread of HIV infection with the regional distribution of cases of malaria imported by Tadjikistan inhabitants. Importation of malaria from other CIS countries was compared as a control. The authors estimated the coefficients of correlation between the affliction of Russia's population with HIV infection and the rate of malaria importation from Tadjikistan (0.733) and other CIS countries (0.496), as well as between the importation of malaria to Russia and the total volume of a migratory flow from Tadjikistan in the period of 1992 to 2001 (-0.931). The findings indirectly support the assumption that there is a relationship of malaria importation from Tadjikistan to illegal migration, which suggests that the intensive importation of malaria from Tadjikistan may serve as a marker of importation of injectable narcotic drugs and hence that the spread of HIV infection in Russia may be intensified. The authors again emphasize that prevention of the spread of socially related infections requires that the whole society should combine efforts since public health care should not be a hostage of social problems. PMID- 15042741 TI - [Enzyme immunoassay of opisthorchiasis in the population of the Shuryshkarsk district, Iamalo-Nenets autonomous region]. PMID- 15042742 TI - [Epidemiological characteristics of pneumocystosis in the Ukraine]. AB - Pneumocysts have been detected in the sputum in patients with nonspecific lung diseases (56 +/- 2%), including those with acute (43 +/- 4%) and chronic (63 +/- 3%) pneumocystosis, in those with a severe course (70.8 +/- 8%); in those with a satisfactory condition (48 +/- 3%); in those with occupational hazards (78 +/- 7%) in smokers (56 +/- 7%); in those without occupational hazards (39 +/- 3%); in those with an inflammation in the upper lobe of the lung in the presence of pneumonia (84 +/- 7%), in cold (62 +/- 3%) and warm (48 +/- 4) seasons, in north (66 +/- 3%) and south (28 +/- 4%) regions. P. carini per ml of sputum numbers in the range of 165 +/- 15 to 195 +/- 25; that in upper lobar pneumonia is 330 +/- 55. In patients with HIV/AIDS, P. carini has been found in the sputum in patients infected with HIV parenterally (66 +/- 4%) and sexually (54 +/- 5%); cysts per ml of sputum number 240 +/- 20 and 200 +/- 20, respectively; the content of CD4+ per microliter of blood is 480 +/- 40 and 205 +/- 30, respectively. PMID- 15042743 TI - [Epidemiological surveillance of parasitic diseases in the republic of Armenia]. AB - Parasitic diseases play a very important role in the regional pathology of Armenia. Malaria was completely eradicated in Armenia in 1963; the malaria-free status was maintained till 1994. At present, the populations of the Ararat, Armavir, and Tavush regions of Armenia are at a potential risk for malaria. Nowadays, 6 species of Anopheles mosquitoes are spreading in Armenia. An analysis of many years' official statistics on the number of individuals infected with intestinal helminthiases and a retrospective analysis of those in 1986-2001 were made to study the current epidemiological laws on intestinal helminthiases. The infection rates of ascariasis, trichocephaliasis, enterobiasis, teniasis, and mixed infection were 4.0 +/- 0.4, 1.2 +/- 0.2, 25.9 +/- 1.0, 0.2 +/- 0.08, and 0.8 +/- 0.2%, respectively. Those of intestinal helminthiasis were higher in rural areas than those in urban areas and in Yerevan. The above fact makes it necessary to work out a new, comprehensive, scientifically-grounded parasitological service adjustable to the present new socioeconomic conditions. PMID- 15042744 TI - [Leptospirosis in Belorussian Polessie: epidemiology and disease prevention]. PMID- 15042745 TI - [Monitoring of the insecticidal resistance of main malaria vectors in Uzbekistan]. AB - In the Fergana valley, the sensitivity of Anopheles maculipennis to DDT was moderate (82.5% death rates) and that to carbamate agents (bendiocarb, 43.5%, and propoxur, 13.0%) was decreased. There was its high sensitivity to pyrethroidal insecticides, such as deltamethrin, lambdacigalothrin, and cifluthrin (100% death rates). The population was moderately resistant to carbamates: bendiocarb and propoxur (69.3 and 77.2% death rates, respectively). In the piedmont and mountainous districts of the Surkhandarya Region, A. superpictus is highly sensitive to all tested drugs. In the plain area of the Samarkand Area, the sensitivity of A. maculipennis to the insecticides, such as fenitrothion, propoxur, and permethrin, was 98.3, 92.3, and 97.0% death rates, respectively; the Tashkent, Dzhizak, and Surkhandaryinsk A. pulcherrimus population retained a normal sensitivity to all tested agents (94.4-100% death rates). The diapausing female A. superpictus mosquitoes in the population from the Fergana valley were found to be resistant to DDT (82.8% death rates) and highly resistant to malathion (43.8%), fenitrothion (38.8%), bendiocarb (2.7%), and propoxur (7.0%); the Tashkent and Surkhandaryinsk populations were also considerably resistant to these agents (35.3-89.6% death rates). PMID- 15042746 TI - [Detection of plague microbe in the fleas by polymerase chain reaction by using magnetic immunosorbents]. AB - The polymerase chain reaction (PCR) with and without pre-treatment of the samples of the fleas Citellophilus tesquorum with magnoimmunosorbents was used to detect the plague microbe in the samples containing 142, 168, 193 or more bacterial cells. PCR analysis reduced the time of tests of ectoparasites for plague to 6 hours. The use of a combination of PCR and magnoimmunosorbent simplified the most time-consuming and longest stage of isolation of plague microbial DNA from the suspensions of the fleas, which allows the time of a test to be reduced to 3 hours. The administration of an affine sorbent having magnetic properties during treatment of samples with plague-infected exoparasites for PCR analysis makes it possible to have a specific concentration of Yersinia pestis and to prevent the inhibitory effect of flea tissues on the polymerase chain reaction, to use the boiling method for isolating microbial DNA and for disinfecting the material to be tested. PMID- 15042747 TI - [Differences between two populations of Citellophilus tesquorum altaicus from the Tuva natural focus of plague]. AB - Citellophilus tesquorum altaicus fleas from two natural (Karginsky and Barlyksky) populations have been studied. There are differences between the fleas of these populations in susceptibility to plague microbe. The adaptability and abundance of the causative organism in the body are higher in the fleas of the Barlyksky population. The sizes of the head are significantly higher in the females of the Karginsky population. There are no differences in this index between males. According to all 5 studies signs of chaetotaxia, the chaeta abundance is greater in both females and males of the Karginsky population. The fluctuating asymmetry is higher in the Barlyksky fleas that are more susceptible to the studied type of the causative agent of plague. PMID- 15042748 TI - [Experimental rationale for the use of medapec as the drug of choice in treating echinococcosis]. AB - The dosage form of medamine-medapec was found to have a high antiechinococcal activity in experiments on laboratory animals. Its efficacy was shown in treating larval alveolar echinococciasis in mice and cotton rats with different doses and courses as compared with medamine and albendazole. It was ascertained that for its high larvicidal activity, medapec should be given to animals regularly during a day. The daily dose of the drug should be gradually increased. In complying with these conditions, the duration of effective courses of therapy drastically reduces. PMID- 15042749 TI - [Activity of the anthelmintic agent trichlorophen on the models of human helminthiasis]. AB - Trials of trichlorophen have shown its high efficacy on models of cestode infections: hymenolepiasis (at the adult and cysticercoid stages of development on three types of animals: outbred albino mice, albino rats and golden hamsters), preimaginal echinococciasis alveolaris, larval alveolar echinococciasis (at the early stage of development of the parasite in experiments on cotton rats). The high nematodical activity of trichlorophen was first found on models of trichocephaliasis in DBA/2y mice, nippostrongyloidiasis (in in vitro experiments), and aspiculuriasis in outbred mice. The agent proved to be ineffective at the tissue developmental stage of Hymenolepsis nana (H. nana), the dwarf tapeworm, in albino mice, during experimental opisthorchiasis in golden hamsters. It showed a low efficacy in treating trichinosis in outbred albino mice. Unlike carbamatebenzimidazoles, trichlorophen was inactive at the tissue stage of H. nana; it exerted no effects on the eggs of a dwarf tapeworm in trichinosis. Trichlorophen was also inactive in treating experimental opisthorchiasis in golden hamsters. PMID- 15042750 TI - [Efficacy of the Russian anthelmintic agent trichlorophen]. AB - Experiments have established the high efficacy of combinations of the micronized dosage form of trichlorophen, with albendazole or medamine in treating trichocephaliasis (its causative agent being Trichocephalus muris) in DBA/2st mice and that of trichlorophen in combination with azinox or fenasal in outbred albino mice with hymenolepiasis (its causative agent being Hymenolepis nana). These combinations are promising in treating patients with cestodosis and nemadosis, respectively. PMID- 15042751 TI - [Synthesis and ovicidal effect of olfelam]. PMID- 15042752 TI - [Strongyloidiasis im Moscow region]. AB - The paper presents two cases of infection with Strongyloides stercoralis (in a 13 year-old boy and in a 93-year-old female), occurred in the Moscow Region, which is indicative of the resurgence of stronglyoidiasis in the Moscow Region. Although the likelihood of this infection in this region is little, nevertheless, strongyloidiasis should be borne in mind in examining patients who are not leaving for well-known endemic areas if they have an allergic reaction of unclear etiology, eosinophilia, and digestive disorders. In these cases, the syndromic and-empiric approach to justifying and performing chemotherapy is recommended due to the inadequate efficiency of parasitological methods for diagnosing strongyloidiasis at the low rate of invasion. These cases are indicative of the rather low virulence of Moscow strains, but at the same time of their capacity for autoinvasion and accordingly for the long-term infection of patients without recurrent infections. PMID- 15042753 TI - [Effect of vitamins on reproductive potential of a population of the ticks Caloglyphus rodionovi (Acariformes: Acaridae) in the laboratory setting]. PMID- 15042754 TI - [Use of a helminthic indicator of soil to determine the frequency of toxoplasmosis among pregnant women]. AB - Studies have ascertained the high dissemination of the soil with helminthic eggs and cysts of intestinal pathogenic protozoa (Lamblia, Toxoplasma, etc.) in some districts of the Belgorod Region. Comparing these data with the morbidity rates in pregnant females with toxoplasmosis shows a direct high correlation of the above indicators. The latter makes it possible to recommend the helminthic indicators of soil contamination to be used to determine the rate of toxoplasmosis in pregnant females. PMID- 15042755 TI - Introduction to the Global Initiative for Asthma and the new guidelines. PMID- 15042756 TI - How to make asthma education happen. PMID- 15042757 TI - "There is more to good asthma care than the prescription alone". PMID- 15042763 TI - [Physiopathology of scarring. III--acute wounds]. PMID- 15042764 TI - [Wound and dressings note. 6--Carbon and silver films]. PMID- 15042765 TI - ["The affair of the veil", a nurse's point of view]. PMID- 15042766 TI - [The right to die?]. PMID- 15042767 TI - [Frequency and risk factors of ORL cancers]. PMID- 15042768 TI - [Diagnosis and therapy of laryngeal cancers]. PMID- 15042769 TI - [Nursing management of the total laryngectomy patient]. PMID- 15042770 TI - [Educational management of the laryngectomy patient]. PMID- 15042771 TI - [Cannula care in the laryngectomy patient]. PMID- 15042772 TI - [During vocal rehabilitation of the laryngectomy patient]. PMID- 15042773 TI - [Giving "speech" back to laryngectomy patients]. PMID- 15042774 TI - [Towards meeting and information places in oncology]. PMID- 15042775 TI - [The Cancer Plan after ten months]. PMID- 15042776 TI - [Cardiovascular system drugs. 1/12 Diuretics]. PMID- 15042777 TI - [1/9. Classification of handicap]. PMID- 15042778 TI - [Psychological care. 4/10 Persons frustrated in the exercise of their role]. PMID- 15042779 TI - [Transcending everyday life]. PMID- 15042780 TI - [Short-term complementary formation in psychiatry?]. PMID- 15042781 TI - [Hospitalization without consent and obligation to care in psychiatry]. PMID- 15042782 TI - [Mediation and nursing care in psychiatry]. PMID- 15042783 TI - [Culture, art and communication in the hospital]. PMID- 15042784 TI - [Establishing an art club]. PMID- 15042785 TI - [Practice of psychology and theater club in the psychiatric service]. PMID- 15042786 TI - [Music therapy club, or when emotions set off]. PMID- 15042787 TI - [Art and therapy: a good encounter]. PMID- 15042788 TI - ["Papers for oneself": a writing workshop]. PMID- 15042789 TI - [Psychoanalysis and family novel]. PMID- 15042790 TI - [Services project and outpatient structures]. PMID- 15042791 TI - What's the proper sales tax? PMID- 15042792 TI - Opinion passing as science. PMID- 15042793 TI - Opinion passing as science. PMID- 15042794 TI - Brian T. Kennedy, D.D.S. The hallmarks of a leader: process, decision and taking responsibility. PMID- 15042795 TI - Role of dental panoramic radiographs in assessment of future dental conditions in patients with osteoporosis and periodontitis. AB - Osteoporosis affects large segments of elderly populations, especially postmenopausal women. Bone mineral density (BMD) assessment of the skeleton by means of several pieces of equipment, such as dual-energy X-ray absorptiometry, have been useful in identifying individuals with low BMD or at high risk of suffering osteoporotic fracture. Recent investigators have demonstrated a significant association between BMD of the mandible and the peripheral skeleton in postmenopausal women. Some studies also have linked low BMD of the mandible and the peripheral skeleton with alveolar bone loss of the mandible and tooth loss. Dental panoramic radiology is a useful imaging modality by which the dentist can evaluate the whole dentition as well as the jawbones. Clinicians have started to focus on some mandibular panoramic indices, such as mandibular cortical index and mandibular cortical thickness, for the identification of elderly individuals who should undergo BMD assessment. In comparison with peripheral BMD measurement equipment in the medical field, the dentist will be able to identify osteoporotic elderly individuals by means of dental panoramic radiographs taken for the diagnosis of the teeth and the jawbones without additional cost. It is likely that the clinician may estimate the future risk of tooth loss in elderly individuals with periodontitis and osteoporosis by dental panoramic radiographs. PMID- 15042796 TI - Diffuse sclerosing osteomyelitis. A case report. AB - Diffuse sclerosing osteomyelitis (DSO) is an interesting and debilitating entity with a disputed, unknown or unproven etiology. This case report illustrates the complexity of managing these cases, which are refractory to multiple therapeutic modalities. PMID- 15042797 TI - Oral health and pregnancy: a review. AB - Pregnancy is a delicate condition involving complex physical and physiological changes. In recent times, there has been a greater focus on the oral health of pregnant women. Emerging evidence has shown that periodontal disease may be associated with preterm, low birthweight (LBW) babies. This has caught the attention of prenatal care providers, who often look to the dental community for information about the oral health aspects of pregnancy. Therefore, dental care providers should be well informed and better able to understand the management of different oral health conditions typically seen during pregnancy. In addition, prenatal care providers should forge a link with oral health care providers to prevent adverse outcomes. This article summarizes the oral health conditions associated with pregnancy. PMID- 15042798 TI - NYU College of Dentistry receives $2 million in NIH funding for osteoporosis research. PMID- 15042799 TI - Influential MSNJ member in N.J. General Assembly. Herbert C. Conaway Jr., MD, JD. Interview by Clark Martin, Patricia A. Costante, and Paul J. Hirsch. PMID- 15042800 TI - Current issues in hormone replacement therapy. AB - Initially used to treat the vasomotor and vaginal symptoms of menopause, hormone replacement therapy (HRT) appeared to have many unexpected beneficial effects in early observational trials. It was hailed as a deterrent of atherosclerosis, osteoporosis, cognitive impairment, and Alzheimer disease. While its salutary effects on bone mass were substantiated, randomized clinical trials noted an increased risk of breast cancer, coronary artery disease, and thromboembolism, and raised doubts about the efficacy of HRT in improving quality of life. This article summarizes the literature and discusses current recommendations on the use of HRT. PMID- 15042801 TI - Clara Louise Maass. The ethics of experimentation on human subjects. PMID- 15042803 TI - [Minimally invasive surgical procedures--the process is going on]. PMID- 15042802 TI - Selecting the right legal counsel. PMID- 15042804 TI - [Endoscopic ultrasound-guided procedures--alternative to surgery]. PMID- 15042805 TI - [Laparoscopic treatment of gallstone disease]. PMID- 15042806 TI - [Abdominal trauma--minimally invasive diagnostics]. PMID- 15042807 TI - [CT and MR colonography]. PMID- 15042808 TI - [Transanal endoscopic microsurgery]. PMID- 15042809 TI - [Laparoscopic surgery of inguinal hernia]. PMID- 15042810 TI - [Laparoscopic treatment of paraesophageal hernias]. PMID- 15042811 TI - [Endovascular treatment of aortic aneurysms]. PMID- 15042812 TI - [Minimally invasive treatment of benign prostatic hyperplasia]. PMID- 15042813 TI - [Robot-assisted urologic and general telesurgery]. PMID- 15042814 TI - [Laparoscopic adrenalectomy]. PMID- 15042815 TI - [Arthroscopic release of shoulder and elbow contracture]. PMID- 15042816 TI - [Female sterilization]. PMID- 15042817 TI - [Video-assisted thoracoscopic treatment of pneumothorax]. PMID- 15042818 TI - [Minimally invasive heart valve surgery]. PMID- 15042819 TI - [Surgery of and local administration to the inner ear]. PMID- 15042820 TI - [Endoscopic treatment of esophago-pharyngeal diverticuli (Zenker's diverticulum)]. PMID- 15042821 TI - [Laparoscopic assessment of resectability of upper gastrointestinal cancer]. PMID- 15042822 TI - [Laparoscopic surgery of colorectal cancer]. PMID- 15042823 TI - [Surgical treatment of uterine bleeding]. PMID- 15042824 TI - [Intralesional treatment of vascular lesions in the head-neck region]. PMID- 15042825 TI - [Picture of the month: vascular ring]. PMID- 15042826 TI - [Hormone therapy and cancer]. PMID- 15042827 TI - [Professional credibility a victim on the altar of ideology]. PMID- 15042828 TI - [Information from the private hospital Hamlet is misleading for the patients]. PMID- 15042829 TI - [Signal noise]. PMID- 15042830 TI - [What do physicians-consultants do in the municipal administration?]. PMID- 15042831 TI - [Microchips based on three dimensional gel cells: history and perspective]. AB - The review describes the history of creation and development of the microchip technology and its role in the human genome project in Russia. The emphasis is placed on the three-dimensional gel-based microchips developed at the Center of Biological Microchips headed by A.D. Mirzabekov since 1988. The gel-based chips of the last generation, IMAGE chips (Immobilized Micro Array of Gel Elements), have a number of advantages over the previous versions. The microchips are manufactured by photo-initiated copolymerization of gel components and immobilized molecules (DNA, proteins, and ligands). This ensures an even distribution of the immobilized probe throughout the microchip gel element with a high yield (about 50% for oligonucleotides). The use of methacrylamide as a main component of the polymerization mixture resulted in a substantial increase of gel porosity without affecting its mechanical strength and stability, which allowed one to work with the DNA fragments of up to 500 nt in length, as well as with rather large protein molecules. At present, the gel-based microchips are widely applied to address different problems. The generic microchips containing a complete set of possible hexanucleotides are used to reveal the DNA motifs binding with different proteins and to study the DNA-protein interactions. The oligonucleotide microchips are a cheap and reliable tool of diagnostics designed for mass application. Biochips have been developed for identification of the tuberculosis pathogen and its antibiotic-resistant forms; for diagnostics of orthopoxviruses, including the smallpox virus; for diagnostics of the anthrax pathogen; and for identification of chromosomal rearrangements in leukemia patients. The protein microchips can be adapted for further use in proteomics. Bacterial and yeast cells were also immobilized in the gel, maintaining their viability, which open a wide potential for creation biosensors on the basis of microchips. PMID- 15042832 TI - [Fluorescence in situ hybridization in studying the human genome]. AB - Physical chromosome mapping by fluorescence in situ hybridization (FISH) is among the major lines of research on the human genome (as well as genomes of numerous other organisms). To localize particular genes or anonymous DNA sequences on individual chromosomes or chromosome regions, FISH was developed in the late 1980s and early 1990s, when the International Human Genome Project and the Russian program Human Genome were launched. Now FISH continues to play a prominent part in studies of the human genome. The review considers the major steps of FISH development in Russia with special emphasis on the key roles of the Institute of Cytology and Genetics (Novosibirsk) and Engelhardt Institute of Molecular Biology (Moscow). Physical mapping of human chromosomes 3 and 13 by FISH is described in detail. The promotion of FISH in Russia contributed to the progress in the related fields such as comparative animal genomics (ZOO-FISH) and studies of plant chromosomes. PMID- 15042833 TI - [Fine structure, expression and polymorphism of the human growth hormone receptor gene]. AB - The human growth hormone receptor gene (GHR) is an example of complex transcription units. The gene has a very long 5'-regulatory region and contains multiple alternative 5'-untranslated exons that are spliced to the protein-coding exons. Its transcription is driven by several promoters that reside far upstream of the coding region. The complete nucleotide sequence of the human GHR gene is known. This paper summarizes the data on the human GHR gene structure analysis and mapping of alternative 5'-untranslated exons. The data of the promoter activity assay in the reporter gene transient expression system, as well as the results of studying the potential regulatory role of the short upstream open reading frame located in one of the 5'-untranslated exons are presented. The search for repeated elements in the human GHR gene was done, and several polymorphic markers that can be used in studies of GHR association with various diseases were characterized. PMID- 15042834 TI - [Molecular-genetic mechanisms of developing the brain based on an embryonic Xenopus model]. AB - The review considers the advantages of Xenopus embryos as an experimental model to study the molecular-genetic mechanisms of embryo development. The results are described that were obtained with this model in studies on the early brain development within the framework of the Russian program Human Genome. PMID- 15042835 TI - [oct-genes and oct-proteins]. AB - The review considers the functions and properties of Oct proteins, which belong to the POU family of transcription factors, and the roles of the POU and other domains in DNA recognition and interaction with other proteins of the transcription initiation complex. The structure and expression regulation of the oct genes are described with special emphasis on alternative transcription initiation from different promoters and alternative splicing, which result in several protein subforms. PMID- 15042836 TI - [Redox-dependent regulation of gene expression induced by nitric oxide]. AB - Nitric oxide (NO) is a mobile, highly reactive signal molecule, and changes the expression of specific genes in effector cells. Under physiological conditions, NO reacts with molecular oxygen and with reactive oxygen species (ROS) to produce intermediates known as reactive nitrogen species (RNS). The production of NO and RNS in the cell is controlled by hormones, neurotransmitters, cytokines, and growth factors. Hence NO and its derivatives act as secondary paracrinous factors and transmit the signal from NO-producing to neighboring cells. Intracellular reception of NO and RNS is due to Src-related tyrosine protein kinases, G-protein Ras, cytochrome oxidase, and guanylate cyclase. Receptor proteins mostly contain heme, active thiol, or iron-sulfur groups, and are both on the plasma membrane and in internal cell compartments. Many of the NO receptors are the key components of cell regulatory systems controlling the transcription factors AP-1, HIF-1, NF-kappa B, and p53 and the expression of their target genes. A distinguishing feature of NO signaling is that changes in redox potential of the cell switch the NO receptor and, consequently, modify the NO effect. Depending on the ROS level, NO activates different signal transduction pathways to induce (or suppress) different gene sets. The data considered indicate that antioxidants may be used to directionally change the transcriptional response of the cell to NO. PMID- 15042837 TI - [Integrated computer system for regulating eukaryotic gene expression]. AB - The review describes several modules of the GeneExpress integrated computer system concerning the regulation of gene expression in eukaryotes. Approaches to the presentation of experimental data in databases are considered. The employment of GeneExpress in computer analysis and modeling of the organization and function of genetic systems is illustrated with examples. GeneExpress is available at http://wwwmgs.bionet.nsc.ru/mgs/gnw/. PMID- 15042838 TI - [Prediction and computer analysis of the exon-intron structure of human genes]. AB - This review of the original works on computer analysis of the human genome considers the development of methods to predict the exon-intron structure of genes and analysis of alternative splicing. Prediction of the gene structure is based on homology between the gene product and a known protein or between the genomic sequences of the gene and its homolog from another organism. The methods were tested and proved highly efficient. Human gene splicing was analyzed with original methods and EST databases. Genes with alternative splicing were for the first time shown to account for no less than 35% total genes. Alternative splicing was compared for the human and mouse genomes. Species-specific isoforms were demonstrated for 50% alternatively spliced genes (25% total genes). PMID- 15042839 TI - [Comparative analysis of primary structure of nucleic acids and proteins]. AB - The review considers the original works on the primary structure of biopolymers, which were carried out from 1983 to 2003. Most works were supported by the Russian program Human Genome and earlier similar Russian programs. Little-known publications of 1983-1993 and recent unpublished results are described in detail. In the field of genome comparisons, these concern the OWEN hierarchic algorithm aligning syntenic regions of two genome sequences. The resulting global alignment is obtained as an ordered chain of local similarities. Alignment of sequences sized about 10(6) nucleotides takes several minutes. The concept of local similarity conflicts is generalized to multiple comparisons. New algorithms aligning protein sequences are described and compared with the Smith-Waterman algorithm, which is now most accurate. The ANCHOR hierarchic algorithm generates alignments of much the same accuracy and is twice as rapid as the Smith-Waterman one. The STRSWer algorithm takes an account of the secondary structures of proteins under study. With the secondary structures predicted using the PSI-PRED software for pairs of proteins having 10-30% similarity, the average accuracy of alignments generated by STRSWer is 15% higher than that achieved with the Smith Waterman algorithm. PMID- 15042840 TI - [System of distributed storage and analysis of genomic information]. AB - A distributed computing system is developed to search and analyze genetic databases using parallel computing technologies. Queries are processed by a local network PC cluster. A universal task and data exchange format is developed for effective query processing. A multilevel hierarchic task batching procedure is elaborated to generate multiple subtasks and distribute them over cluster units under dynamic priority levels and with dynamic distribution of replicated source data subbases. Primary source data preparation and generation of annotation word indices are used to significantly reduce query processing time. PMID- 15042841 TI - [Genomic and molecular medicine]. AB - Molecular medicine is a new research field underlain by achievements of the Human Genome Project. The review considers the contribution of the Laboratory of Prenatal Diagnostics of the Ott Institute of Obstetrics and Gynecology to the development of molecular medicine in Russia. Special emphasis is placed on molecular diagnostics, predictive medicine, and gene therapy. The lab obtained priority results in devising and promoting methods of molecular diagnostics of the most common severe hereditary disorders such as cystic fibrosis, Duchenne muscular dystrophy, hemophilia A, and fragile X syndrome. Owing to the Russian program Human Genome, St. Petersburg researchers laid the foundations for theoretical and applied predictive medicine, which is aimed at identifying and analyzing the genes associated with predisposition to high-incidence multifactorial disorders. Experiments with mdx mice providing a model of Duchenne muscular dystrophy were carried out to select the optimal way of delivering a transgene (cDNA of the dystrophin gene) contained in various constructs for the purpose of gene therapy. PMID- 15042842 TI - [Human molecular genetics: study in the area of medical and ethnic genomics]. AB - The review covers selected research topics in fields of medical and ethnic genomics tackled at the Department of Molecular Basis of Human Genetics, the Institute of Molecular Genetics. Primary concern is given to genetic causes of monogenic neurological disorders, among them hepatolenticular degeneration (Wilson's disease), torsion dystonia, and myotonic dystrophy. Results of polymorphic DNA marker surveys in Russia and neighboring countries are also presented. PMID- 15042843 TI - [Genomic study of the hereditary pathology and genetic diversity of the Siberian population]. AB - The review considers the results of genome research on the Russian program Human Genome carried out in the Institute of Medical Genetics (Tomsk) from 1990. The three major fields were molecular cytogenetics and chromosomal disorders, genomics of Mendelian and high-incidence diseases, and ethnogenomics of the North Asian population. Several human genes were cytogenetically mapped, and numerical and structural abnormalities associated with human diseases studied by fluorescence hybridization. Procedures of DNA diagnostics were developed for 15 hereditary diseases. New data were obtained on genetic heterogeneity of idiopathic hypertrophic cardiomyopathy. The genetic bases of multifactorial (atopic bronchial asthma) and infectious (tuberculosis) diseases were analyzed. The North Eurasian population (41 local populations of 21 ethnic groups) was tested for genetic diversity with numerous genetic markers, including Y chromosomal haplotypes, autosomal microsatellites, and polymorphic Alu insertions. PMID- 15042844 TI - [Genomic structure and DNA diagnosis of hereditary monogenic diseases in the Volga-Ural region]. AB - The review considers the main results of molecular analysis of the genes responsible for cystic fibrosis, phenylketonuria, Wilson-Konovalov disease, Duchenne-Becker progressive muscular dystrophy, myotonic dystrophy, Huntington's disease, and nonsyndromic hereditary hypoacusis in populations of the Volga-Ural region. The results were obtained in the past ten years within the framework of the Russian program Human Genome. The mutation spectra and frequencies of these genes were characterized in the major ethnic groups (Bashkirs, Tatars, Russians) of Bashkortostan. Several diseases were associated with particular alleles or haplotypes of polymorphic loci of relevant genes. The results were used to develop DNA diagnostic procedures optimal for the region and to establish the origin of the mutations involved. PMID- 15042845 TI - [Genomics of type I diabetes mellitus and its late complications]. AB - In ethnic Russians, MHC (HLA) was shown to be the major locus determining the predisposition to type 1 diabetes mellitus (T1DM). To map the regions linked to T1DM, families with concordant or discordant sib pairs were selected from the Russian population of Moscow. With these families, linkage to T1DM was demonstrated for CTLA4 (IDDM12, 2q32.1-q33), which codes for a T-cell surface antigen, and PDCD2 (IDDM8, 6q25-q27), which is homologous to the mouse programmed cell death activator gene. With polymorphic microsatellites, regions 3q21-q25 (IDDM9) and 10p12.2 (IDDM10) were also linked to T1DM. Case/control and family studies of the polymorphic markers from region 11p13 revealed a new T1DM associated locus in the vicinity of the catalase gene (CAT); linkage to this locus was not reported earlier for other populations. Diabetic polyneuropathy (DPN) proved to be associated with single-nucleotide polymorphisms Ala(-9)Val (SOD2), Arg213Gly (SOD3), and T(-262)C (CAT) and with a polymorphic microsatellite of the NOS2 promoter. Hence oxidative stress, which results from hyperglycemia, increased mitochondrial production of superoxide radicals, and insufficient activities of antioxidative enzymes, was assumed to play an important part in DPN development in T1DM. Diabetic nephropathy (DN) showed no association with the antioxidative enzyme genes. However, the association was observed for the insertion/deletion (I/D) polymorphism of ACE and the ecNOS34a/4b polymorphism of NOS3, two genes involved in controlling vascular tonicity, and for the I/D polymorphism of APOB and the epsilon 2/epsilon 3/epsilon 4 polymorphism of APOE, two genes involved in lipid transport. In addition, polymorphic microsatellites of chromosome 3q21-q25 proved to be closely associated with DN. The tightest association was established for D3S1550, carriers of allele 12 or genotype 12/14 having high risk of DN (OR = 4.85 and 6.25, respectively). Region 3q21-q25 was assumed to contain a major gene determining DN development, while the other DN-associated genes mostly affect the progression of DN. PMID- 15042846 TI - [Genomics in psychology and psychiatry]. AB - The review considers the most interesting data on the molecular genetic basis of mental disorders and personality traits, which were obtained within the framework of the Russian program Human Genome. Polymorphic markers Taq1A of the dopamine receptor gene and T102C of the serotonin receptor type 2A gene were associated with schizophrenia, in particular, chronic forms with poor prognosis. In mentally healthy people, the insertion/deletion polymorphism of the serotonin transporter gene was associated with schizoid traits. PMID- 15042847 TI - [The main tasks of Czech perinatology at the beginning of the 21st century]. AB - OBJECTIVE: To identify the main goals of the Czech perinatology at the beginning of the 21st century. DESIGN: Retrospective epidemiologic analysis of selected perinatal care quality indicators. SETTING: Institute for the Care of Mother and Child, Prague. METHODS: Analysis of selected indicators of perinatal care and their trends in the Czech Republic and comparison with other countries. RESULTS: When we reached the same low level of perinatal mortality as the other most developed countries, we have observed unchanged values for last three years. The priorities established 15 years ago are still important to preserve the current situation, however the further development in perinatology requires now goals related to development of selected indicators of perinatal care and their international comparisons. These are: 1) change of definition of stillbirth including newborns of 500-999 grams (currently registered as spontaneous abortions), 2) termination of the continuous increase of Caesarean section, 3) inclusion of the criteria of quality of perinatal care, especially a) deaths during late neonatal and postneonatal periods with perinatal causes; b) perinatal morbidity (impaired neuro-sensoric and psychological development of the child due to perinatal causes). Both these tasks will require collaboration of professionals from obstetrics and neonatology with specialists from other fields. CONCLUSIONS: The 4 main tasks were established on the basis of development of selected perinatal care quality indicators and their international comparisons: the change of definition of stillbirth, the termination of continuous increase of Caesarean sections, and inclusion of other indicators of perinatal care, first related to mortality after 7th day of life due to perinatal causes, and second the perinatal period related morbidity in later periods of infant life. PMID- 15042848 TI - [Psychological aspects in perinatal medicine 1980-2000: III. The postnatal period]. AB - OBJECTIVE: Analysis of the problems concerning postnatal period in the Czech Republic over the last 20 years from psychological point of view. The analysis is included in the basic materials for the elaboration of prognosis in the given area for the project called "Evaluation of the evolution, present state and conditions to secure healthy physical, mental and social development of the new generation until the year 2010" (Grant No. 7346-3). DESIGN: A retrospective analysis of the problems concerned of the postnatal period from the viewpoint of the psychologist in the Czech Republic in the period of 1980 to 2000. SETTINGS: Institute for the Care of Mother and Child, Prague. CONCLUSIONS: In the postnatal period great changes have occurred over the last 20 years especially in connection with introduction of a joint care of the mother and the newborn by so called "rooming-in" system. This kind of system, which favorably influences the relations between mother and child as well as the breast-feeding, is mostly accepted favorably by the women. In the Czech Republic, low attention has been paid so far to postnatal psychical disorder in the women, especially to so called post partum or postnatal depression, which occurs in 10 to 15% of puerperas and exerts unfavorable effect on the formed relationship between the mother and child. It should be useful to contact these women in time and offer them a sufficient psychosocial support. The psychosocial care of the newborns at risk and their parents in our society requires to keep up with the advanced western countries, as well as the care of parents whose newborn dies. PMID- 15042849 TI - [Analysis of congenital defects contributing to perinatal mortality in the Czech Republic]. AB - OBJECTIVE: Analysis of the proportion of selected types of congenital defects in stillbirth, frequent neonatal and perinatal death rate in the Czech Republic in the years 1994-2002. DESIGN: A retrospective demographic-epidemiological analysis. SETTING: Institute for the Care of Mother and Child, Prague. MATERIALS AND METHODS: The study analyzed individual anonymous data from the Institute of Medical Information and Statistics of the Czech Republic in the period of 1994 2002 (preliminary data only for 2002) from the National Registry of Congental Defects. Fourteen types of defects were selected from the data set. The proportion of children with a certain selected congenital defect which died during the perinatal period were calculated in comparison with the total number of subjects diagnosed for this congential defect in the Czech Republic over the period of observation. RESULTS: A total of 4,436 children died during the perinatal period in the Czech Republic over the period of 1994-2002, while 879 of them suffered from one or more congenital defects. The congenital defects represent more than 10% of stillbirth in this period, 32.5% of deceased individuals in early neonatal death rate and almost 20% in perinatal death rate. In 25% of these cases the prenatal diagnosis was positive, whereas prenatal diagnosis was negative or not indicated (a negative screening) in 48% of cases, and in 27% the data were not filled out. More than 45% of children deceased with a congenital defect in the perinatal period had two or more congenital defects. CONCLUSIONS: Analysis of these results from the registry of congenital defects in the Czech Republic contributed to the information on prenatal deaths in the period of 1994-2002. The proportion of congenital defects in perinatal death rate makes up almost 20%, but it also significantly participates in late newborn and post-newborn mortality and morbidity. PMID- 15042850 TI - [Role of prenatal diagnosis in decreasing the incidence of congenital defects in the Czech Republic]. AB - OBJECTIVE: An analysis of prenatal diagnostics of congenital malformations in the year 2002 and an analysis of the proportion of prenatal diagnostics in the total incidence of congenital malformations in the Czech Republic in the years 1994 2002. DESIGN: A retrospective study. SETTING: Department of Medical Genetics, Faculty Thomayer Hospital, Prague. MATERIALS AND METHODS: The data were obtained from Dept. of Medical Genetics wards (28 departments)--congenital malformations were detected during prenatal diagnostics, and other data were obtained from the Institute of Medical Information and Statistics of the Czech Republic, the National Registry of Inborn Defects--Congenital Malformations in Newborns. From the available material the authors selected six types of congenital malformations (anencephaly, spina bifida, omphalocoele, gastroschisis, renal agenesis/hypoplasia, Down syndrome) for a more detailed analysis. RESULTS: In the period of 1994-2002 a total of 3,621 pregnancies with congenital malformations were detected and terminated and 1,351 fetuses were prenatally diagnosed, but the pregnancy was not terminated. The prenatal detection of inborn errors is gradually increasing. A detailed analysis of prenatal diagnostics in 2002 indicated that chromosomal aberrations were diagnosed most frequently (307 cases), of which Down syndrome was most frequent (108 cases). The next most frequent defects were those of urinary tract (agenesis or renal hypoplasia 13 times, kidney cystosis 25 times, hydronephrosis 31 times, other congenital malformations 10 times), congenital heart malformations (dealt with in another presentation), defects of neural tube (anencephaly 18 times, encephalocoele 6 times, spina bifida 22 times), congenital malformations of skeletal and muscular system (46 times) and defects of abdominal wall (omphalocoele 10 times, gastoschisis 25 times). A detailed analysis of six types of congenital malformations revealed that prenatal detection of congenital malformations increases with time and prenatal diagnostics significantly participates in the total detection of congenital malformations. The percentage of prematurely terminated pregnancies from the total number of diagnosed malformations is almost 100% in anencephaly, 50-60% in spina bifida, 50-60% in omphalocoele, 70-90% in gastroschisis, 20-30% in renal agenesis/hypoplasia and 60-70% in Down syndrome. CONCLUSION: In the period of observation there were increasing numbers of prenatally diagnosed congenital malformations. A difference develops between the total number of prenatally diagnosed cases and those diagnosed prenatally and terminated. There is an increasing number of non-terminated cases (decision of the pregnant woman, multiple pregnancies, late detection of the congenital malformations). The efficiency of prenatal diagnosis in the observed period was in the range of 50-100% in relation to the type and severity of the congenital malformations. PMID- 15042851 TI - [Incidence of congenital defects in the children of mothers who used medications in the first trimester of pregnancy in the Czech Republic 1996-2001]. AB - OBJECTIVE: Analysis of the results from registration of congenital defects in children of mothers taking therapeutic drugs in the first trimester of pregnancy and comparison with a control group in the Czech Republic in the period of 1996 2001. DESIGN: A retrospective analysis of data from the registry of congenital defects in the Czech Republic. SETTING: Institute for the Care of Mother and Child, Prague. METHODS: Analysis of incidences of selected types of congenital defects detected in newborns of mothers taking therapeutic drugs during the first three months of pregnancy. The paper employed data from the all-state registration of congenital defects held in the Institute of Medical Information and Statistics of the Czech Republic in the period of 1996-2001. Data from healthy children born to mothers who took therapeutic drugs during the first trimester of pregnancy were used as a control set. RESULTS: In the period we observed, a total of 17,674 cases of children with diagnosed congenital defect were detected in the registration of congenital defects in the Czech Republic. In this number there were 784 cases of children whose mothers used therapeutic drugs during the first trimester of pregnancy. The control group included 1,034 women who gave birth to healthy children without congenital defects, although they took therapeutic drugs during the first trimester of pregnancy. Statistically significantly higher risk was found in 13 groups of diagnoses: anencephaly, inborn hydrocephalus, spina bifida, inborn defects of eyelids, lacrimal system and orbita, anoftalmus, microphthalmus and macrophthalmus, inborn defects of ear, congenital defects of the heart septum, congenital defects of great veins, cleft palate with cleft lip, congenital defect of gall bladder, biliary pathways and liver, congenital deformities of the hip, reduction deformities of upper extremity, congenital defects of muscular and skeletal system. Significantly higher risks were found for five groups of therapeutic drugs: anticoagulants, antihypertensive drugs, peripheral vasodilatants, urological drugs and antiepileptic drugs. CONCLUSION: It is obviously impossible to draw significant conclusions with clinical consequences on the basis of these results. Nevertheless, our results supplemented international databases of untoward effects of drugs and the conclusions may become a part of data set necessary for analysis of possible teratogenic effects of drugs used in the critical developmental period during the first trimester. The pregnant women should avoid, during the first trimester, all drugs except those, which are carefully medically indicated and accepted as adequately safe. The administration of other drugs should be evaluated by a clinical geneticist, who should take into account genetic and teratogenic risk in individual cases. PMID- 15042852 TI - [Undifferentiated choriocarcinoma--epithelioid trophoblastic tumors treated at the Center for Trophoblastic Diseases in the Czech Republic 1955-2003]. AB - OBJECTIVE: The clinical-pathological picture, pathogenesis, biological behavior and therapy of epithelioid trophoblastic tumor (ETT) alias undifferentiated choriocarcinoma (CH-Ned). DESIGN: A retrospective analysis. SETTING: Trophoblastic Disease Center in the Czech Republic (TDC-CZ), Department of Gynecology and Obstetrics, 3rd Medical Faculty, Charles University, Institute for the Care of Mother and Child, Prague. METHODS: The identification of all tumors complying with histopathological criteria of ETT-CHNed among 372 malignant tumors of trophoblast (MTT), treated at TDC-CZ in the years 1955-2003. Their morphological analysis was done from the standpoint of formal pathogenesis, correlation with clinical picture, laboratory and therapeutic results. RESULTS: Among 372 malignant tumors of trophoblast (MTT) we detected 25 ETT-CHNed. The size of the tumor was in the range of 15 to 45 mm except two cases. One tumor diffusely infiltrated thyroid gland and clinically imitated struma. In the other case a massive dissemination of ETT-CHNed in the lungs was supposed to be tuberculosis. The tumor in the uterus and metastases was predominantly of solid character, not infrequently with necroses and haemorrhages. The microscopic analysis revealed larger irregular cells with frequent mitoses, resembling eight day orthologic trophoblast. There were also infrequent elements of cyto intermediate and syncytium-trophoblastic character. The mitotic index (3-7), proliferation markers (20%), inhibitin alpha, hCG and PLAP with histological picture suggest a specific form of MTT or choriocarcinoma. The age of the female patients was in the range of 22 to 43 years. In 18 cases (72%) the tumor displayed gynecological symptomatology, in 7 cases (28%) a non-gynecological one (pulmonary 3 times, thyroid once, CNS once, GIT once, mamma once). In the case history there was delivery in 10 cases, abortion in eight, mola hydatiosa completa twice, anamnesis was uncertain once and extra-uterine pregnancy was suspected also once. The interval between pregnancy and established diagnosis was in the range of one to 64 months. The ETT-CHNed diagnosis was established 18 times from curettage of endometrium, six times from biopsies of organs considered as primary localization of the tumor and once during post mortem examination. The hCG values upon admission were in the range of 10(1) to 10(3) mIU/ml in connection with a small number of differentiated syncitium-trophoblastic cells. In the first period (1955-1963) before introduction of chemotherapy all five patients died (100%) in the range of 4 months to 3 years. In the second period (1964-1980), hysterectomy with subsequent monochemotherapy resulted in permanent remission (20 years) in four women out of nine (44%). In the third period (1981 2003), hysterectomy with subsequent polychemotherapy resulted in complete remission from two to 18 years in 9 out of 11 women (82%), while in two cases with absent ETT-CHNed in uterus the intervention was limited to tumor extirpation in the lung or mamma with subsequent treatment with chemotherapy. CONCLUSION: The revision of 372 MTT treated in TDC-CZ (1955-2003) uncovered 25 ETT-CHNed. Their clinical-pathological analysis revealed that ETT-CHNed is a malignant tumor, which is not less aggressive than choriocarcinoma (CH-NST). It becomes manifest by frequent metastases, often with absent demonstrable tumor in the uterus. It represents a less differentiated form of MTT, becoming manifest in a low production of hCG. It can be derived by formal pathogenetics from non differentiated earliest orthologic trophoblast. There are differentiated transitions between ETT-CHNed and CH-NST, which are analogous to grading of other malignant epithelial tumors. Hysterectomy with subsequent intensive chemotherapy decreased the original 100% mortality in the years 1955-1963 to 18.1% in the years 1980-2003. PMID- 15042853 TI - [Lymphangiogenesis and new markers for lymphatic endothelium]. AB - OBJECTIVE: To summarise recent knowledge about lymphangiogenesis, markers of the lymphatic endothelium and their importance to the tumor dissemination. DESIGN: A literature review. SETTING: Department of Gynecology and Obstetrics, Medical Faculty, Palacky University, University Hospital, Olomouc. ABSTRACTS: Clinicopathological data suggest that the lymphatics are an initial route for the spread of solid tumors. Detection of sentinel lymph nodes by biopsy provides significant information for staging and designing therapeutic regimens. The role of angiogenesis (formation of new blood vessels) in facilitating the growth of tumors has been well established, but the presence of lymphatic vessels and the relevance of lymphangiogenesis (formation of new lymphatics) to tumor spread are less clear. Growth factors involved in lymphangiogenesis during embryogenesis have been recently identified and showed to be able to induce formation of intratumoral lymphatics and thus facilitate metastatic spread to lymph nodes. The development of approaches to block tumor lymphangiogenesis and treat lymphedema would benefit from the availability of markers specific for lymphatic endothelium. PMID- 15042854 TI - [Host-yeast interactions in relation to the urogenital tract, vulvovaginal candidiasis and urologic aspects of mycotic diseases]. AB - OBJECTIVE: A review of the role of yeasts in indigenous microbiota with the focus on urogenital tract. The yeast in vaginal environment and their relation to acute and recurrent form of vulvovaginal candidiasis. DESIGN: A review. SETTING: Department of Obstetrics and Gynecology, Department of Urology, Medical Faculty, Charles University, Hradec Kralove. SUBJECT AND METHODS: Review of current literature data with the contribution on own experience. CONCLUSIONS: Vulvovaginal candidiasis is one of the most frequent complaints in patients attending gynecologist. Recurrent form of this disease is still an unsolved problem because the etiopathogenesis is unexplained and successful therapeutic management from the point of view of next attacks is very difficulty. PMID- 15042855 TI - [Treatment of aerobic vaginitis and clinically uncertain causes of vulvovaginal discomfort]. AB - OBJECTIVE: The treatment of clinically uncertain conditions of vaginal discomforts with a mixed preparation of nifuratel + nystatin (Macmiror complex) and the relation of uncertain conditions to aerobic vaginitis. DESIGN: A prospective study. SETTING: Gynecology-Obstetrics Outpatient Department LEVRET Ltd., AescuLab Ltd., Laboratory of Microbiology, Prague. METHODS: 50 women with vaginal discomfort, causes of which had not been clarified by gynecological examination, determination of pH and the amine test, were examined by vaginal smears using microscopy. The results were evaluated in relation to aerobic vaginitis in a pure form or in combination with other nosological units. The authors also evaluated results of therapy by oral nifuratel (Macmiror tbl) 3 x 200 mg daily and a vaginal combined preparation containing nifuratel 500 mg + nystatin 200 kIU (Macmiror complex 500 glo vag) for the period of 7 days. RESULTS: In 50 women candida was demonstrated 24 times, presence of key cells 11 times, lactobacillus nine times with more than 50 in the field, six women were affected by aerobic vaginitis. In all these cases the pH was 4.8 or higher, leukocytes were significantly represented in all cases (> 15 in the field), as well as gram-negative bacteria and/or cocci (> 30 in the field), indicating a combined picture of mycosis, anaerobic vaginosis or lactobacillosis with aerobic vaginitis. The therapy was successful in all cases, the relapse of complaints during one month occurred in three cases. CONCLUSIONS: Aerobic vaginitis in a pure form or with anaerobic vaginosis, mycosis or lactobacillosis is frequently concealed under clinically uncertain pictures of vulvo-vaginal discomfort. The therapy by a combination of nifurated 3 x 200 mg orally together with the combined vaginal preparation nifuratel 500 mg + nystatin 200 kIU for the period of 7 days exerts high effect and a low number of relapses. PMID- 15042856 TI - [DNA analysis in gestational trophoblastic disease]. AB - OBJECTIVE: DNA analysis of different forms of gestational trophoblastic disease. DESIGN: Retrospective clinical study. SETTING: Slovak Center of Trophoblastic Disease, Bratislava, Slovak Republic. METHODS: In the period of September 1993 to April 2003, eighty-nine cases of gestational trophoblastic disease were analysed. There were 22 cases of partial hydatidiform moles, 58 cases of complete hydatidiform mole, 5 cases of invasive mole and 4 cases of gestational choriocarcinomas. Southern hybridization and polymerase chain reaction were used for DNA analysis. RESULTS: From 22 analyzed cases of partial hydatidiform moles 19 (86.4%) were triploid and 3 (13.6%) diploid ones. There were 58 cases of complete hydatidiform mole and out of them 29 (50%) were homozygous, 28 (48.3%) heterozygous, and in one case (1.7%) both paternal and maternal genome was detected. In 8 cases of heterozygous and in one case of homozygous complete hydatidiform mole occurred a malignant transformation to gestational choriocarcinoma. CONCLUSIONS: Molecular analysis can determine the nuclear DNA origin of complete hydatidiform mole and allow us to define the patients with higher risk of malignant transformation usually to gestational choriocarcinoma. PMID- 15042857 TI - [Early cleavage embryos and their effect on the results of standard in vitro fertilization]. AB - OBJECTIVE: To assess if early cleavage embryo is an additional indicator of embryo quality that could be used to increase the efficiency of IVF and ET. DESIGN: A prospective study. SETTING: Centre for Assisted Reproduction, Department of Gynecology and Obstetrics, Medical Faculty, Palacky University, University Hospital, Olomouc. METHODS: The study evaluates 62 cycles of conventional IVF in our Centre during 2001 to 2002. All embryos were observed for the presence or absence of "early cleavage" (EC), i.e. with completed first mitotic division 25 to 26 hours after insemination. The EC embryos were cultivated separately from the so called "late cleavage" or no early cleavage (NEC) embryos. After the three-day cultivation a maximum of three embryos were transferred in 6-8 cells stage. EC embryos were transferred in 28 cycles, NEC embryos in 18 cycles. The remaining 16 patients with "combined" transfer of EC + NEC embryos were excluded from the evaluation. The following parameters have been compared: age, number of fertilized oocytes, fertilization rate (FR), number of cryopreserved embryos, number of embryos assessed for EC, number of EC embryos found, number of transferred embryos, pregnancy rate (PR), implantation rate (IR), spontaneous abortions (AB). Statistical analysis was performed by Student's t-test, chi 2 test and Sheffe analysis. RESULTS: EC embryos were found in 71% of conventional IVF (n = 44). There were no significant differences in the entry parameters (age, number of fertilized and fertilizing oocytes, number of cryopreserved, assessed and transferred embryos) between the group with EC embryos versus the group with NEC embryos. When EC embryos were transferred, the following parameters were higher: PR (61% vs. 56%), IR (38% vs. 33%), compared to NEC group, while pregnancy loss was lower after EC (4% vs. 11%) compared to NEC. Due to small numbers in the subgroups the results, though suggestive, are not statistically significant (p > 0.05). CONCLUSIONS: The assessment of early cleavage embryo is one of the criteria for evaluation and selection of suitable embryos prior to transfer which could help in the process of selection to choose the best embryos between embryos of the same morphology on the day of transfer. PMID- 15042858 TI - [Successful treatment of azoospermia diagnosed as "Sertoli cells only syndrome with maturation arrest"--developing a quality embryo]. AB - OBJECTIVE: The transfer of good quality embryo in the program of assisted reproduction in the case of azoospermia, dg. Sertolli cells only syndrome (SCO sy) + maturation arrest (MA). Testes were assessed and found to have a high occurrence of Sertolli cells and very low occurrence of germinal cells, which were arrested at the round spermatid level. The histological evaluation was hypospermatogenesis gr. 3 (minimum 1 spermatid/sample). DESIGN: Case report. SETTING: Laboratory IVF, Iscare, a. s., Department of Biology and Biochemistry of Fertilization, Institute of Molecular Genetics, Czech Academy of Sciences, Prague. SUBJECT AND METHOD: The successful integration of three methods provides a solution for this case of azoospermia. Immunology and histology can more exactly diagnose the degree of azoospermia. Detection and visualisation of spermatids using monoclonal antibodies against sperm proteins predicts the eventual occurrence of spermatogenesis, and histological evaluation confirms these immunological findings. Using the information of both methods it is possible to use special in vitro cultivation of testicular cells and so obtain injectable spermatozoa, or precursors of sperm, for the ICSI method. CONCLUSION: The probability of acquisition of good-quality embryo in the program of assisted reproduction is higher when these three methods are applied in combination. PMID- 15042860 TI - Proposed 2004 amendments to ONA bylaws. PMID- 15042859 TI - Vulvar rhabdomyosarcoma in a 15-year-old girl (case report). AB - Knowledge of vulvar rhabdomyosarcoma (RMS) is based on case reports and collective reviews since the pathology is extremely rare and experience of individual institutions limited. We present a case of a 15-year-old girl with large-sized vulvar embryonal RMS. The patient was treated with wide local excision of the tumor, multiagent chemotherapy and radiotherapy, achieved complete remission but 6 months after diagnosis died of toxicity. The management is discussed. Necessity of early anticipation of malignancy in girls is pointed out as it may determine outcome of therapy. PMID- 15042861 TI - Inpatient management courses at ONA. PMID- 15042862 TI - ANA adopts precautionary principle. PMID- 15042863 TI - [The role of ventromedial prefrontal cortex in addiction disorders]. AB - Recently a significant conceptual change emerged in the interpretation of addictological disorders. Despite this significant progress, the exact neuronal mechanisms of these disorders are still unknown. By the development of cognitive neuroscience novel clinical tests became available which are devoted to the evaluation of more properly defined neuronal structures. These novel approaches are designed to separately investigate memory/attention-related and affective processes in decision-making. These investigations along with animal models and functional neuroimaging approaches suggest the crucial role of a complex neuronal network in the pathomechanism of addictological disorders, including the ventromedial prefrontal cortex and the amygdala-accumbens system. The aim of this paper is to review these novel findings in order to gain insight into the neurocognitive mechanisms of addictological disorders. PMID- 15042864 TI - [Clinical features and diagnosis of spinocerebellar ataxia]. AB - In the past decade, a great progress has been made in understanding genetic basis of the spinocerebellar ataxia. Based upon the genotypes, more then 20 subgroups of autosomal dominant spinocerebellar ataxia have been identified with different gene mutations. Neither the pathomechanism nor the function of these genes is fully understood. In these disorders the main clinical sign is ataxia. Other symptoms may be present as well, but no specific clinical feature is known for differentiating subgroups. Specific diagnosis can be made by genetic tests. In this review we summarize the clinical features and genetic backgrounds of the most common spinocerebellar ataxias. PMID- 15042865 TI - [Effect of mental arithmetic and verbal fluency on blood flow velocity in the middle cerebral arteries]. AB - INTRODUCTION: Using transcranial Doppler sonography (TCD), changes in blood flow velocity (BFV) can be measured in the Medial Cerebral Artery (MCA) during cognitive effort. Our goal was to define the time-course and laterality of BFV in healthy volunteers during arithmetic and verbal fluency tasks according to handedness. METHODS: Twelve subjects (8 right-handed, 4 left-handed) were assessed. The TCD registered BFV in both MCA simultaneously. Heart rate was also recorded using TCD. Finally we included a 16-channel EEG. BFV laterality index (LI) was calculated. Participants were asked to count silently and generate words beginning with a specified letter. To estimate hemispheric differences in BFV, two-tailed Wilcoxon tests were utilized along with correlational analyses. RESULTS: During cognitive effort the BFV changed in a tri-phasic manner in all participants. A 6-8% elevation of BFV was observed in MCAs without latency at the time of the evoking signal. Laterality of BFV developed after 5-13 seconds during cognitive effort in right-, and several seconds later in left-handed subjects. During tasks the BFV increased in the dominant hemisphere up to 2.6-4.7% compared to the subdominant one. We also calculated the LI. During the verbal task the LI agreed with the handedness in 9 out of 12 subjects. During the mental arithmetic task, agreement was found in 6 out of 12 subjects. According to LI results we found a discrepancy between verbal and arithmetic tests in 3 out of 12 subjects. CONCLUSION: Cognitive effort elicites significant bilateral BFV increases in the MCAs, which suggests fast neurogenic regulation. The course of BFV during mental arithmetic proved to be different from course BFV assessed during the word fluency task. Based on the laterality of the BFV, the word-generation task was more sensitive in determining the dominant hemisphere when compared to the mental arithmetic task. The use of LI may help to estimate hemispheric functions even in pathologic circumstances. PMID- 15042866 TI - [Brachytherapy of brainstem tumors]. AB - INTRODUCTIONS: The optimal therapy of brain stem tumours of different histopathology determines the expected length of survival. Authors report 125Iodine interstitial irradiation of brain stem tumours with stereotactic brachytherapy. CASE REPORTS: Two patients having brain stem tumours were suffering from glioma or from metastases of a carcinoma. In Case 1 the tumour volume was 1.98 cm3 at the time of planning interstitial irradiation. The control MRI examination performed at 42 months post-op showed a postirradiation cyst size of 5.73 cm3 indicating 65.5% shrinkage. In Case 2 the shrinkage was more apparent as the tumour volume measured on the control MRI at 8 months post-op was only 0.16 cm3 indicating 97.4% shrinkage of the 6.05 cm3 target volume at the time of brachytherapy with the metastasis practically disappearing. Quick access to histopathological results of the stereotactic intraoperative biopsy made it possible to carry out the 125Iodine stereotactic brachytherapy immediately after the biopsy, resulting in less inconvenience for patients of a second possible intervention. The control MRI scans show significant shrinkage of tumours in both patients. CONCLUSION: The procedure can be performed as a biopsy. The CT and image fusion guided 125Iodine stereotactic brachytherapy can be well planned dosimetrically and is surgically precise. PMID- 15042867 TI - [A novel analgesics made from Cannabis]. AB - Bayer AG has recently announced that it acquired exclusive rights for the marketing of GW Pharmaceuticals' new medicine Sativex in Europe and in other regions. Sativex is a sublingual spray on Cannabis extract basis, and is equipped with an electronic tool to facilitate accurate dosing and to prevent misuses. It is standardized for the THC and CBD. The new analgesic is proposed for the treatment of muscle spasticity and pains accompanying multiple sclerosis and as an efficient analgetic for neurogenic pain not responding well to opioids and to other therapies available. The entirely new mechanism of action through the recently discovered cannabinoid receptor system may offer a real therapeutic potential to the drug. Although the Government of Netherlands has authorized the sale of pharmaceutical grade Cannabis herb by pharmacies in the Netherlands, the availability on the pharmaceutical market of the registered preparation may render requests for the authorization of the smoking of Cannabis herb (marihuana) by individuals suffering of multiple sclerosis, neurogenic pain, AIDS wasting syndrome unnecessary. Nevertheless, the "old chameleon" plant Cannabis appears to gradually regain its previous status in mainstream therapy and pharmacy. As long as the plant Cannabis and its products continue to be classified as narcotic drugs, medical use of the new preparation will need close supervision. PMID- 15042868 TI - Multiple sclerosis to date: diagnosis, epidemiology, new aspects of the pathomechanism and the therapy. PMID- 15042869 TI - Outcomes of Christian nursing. PMID- 15042870 TI - Second chance. Seizing a sacred moment. PMID- 15042871 TI - The baptism of Grady Flynn. PMID- 15042872 TI - How are we doing? Measuring the effectiveness of parish nursing. PMID- 15042873 TI - How well do parish nurses document? PMID- 15042874 TI - Church nurse. Building on a proud tradition. PMID- 15042875 TI - Journey into hope. A therapeutic relationship. PMID- 15042876 TI - Do unto others ... when you'd rather not. PMID- 15042878 TI - I see glory! PMID- 15042877 TI - Does spiritual care really help? A study of patient perceptions. PMID- 15042879 TI - Unexpected outcome. PMID- 15042880 TI - Responding to God's call. Outcomes of faith. PMID- 15042881 TI - Dental amalgam and mercury toxicity: should dentists be concerned? PMID- 15042882 TI - Aesthetic restoration of the compromised root: a case report. AB - The amount of tooth structure that remains after endodontic therapy and post preparation is of prime importance as the strength and resistance to root fracture of an endodontically treated tooth is directly related to the amount of residual dentine. The desired post diameter is determined by tooth morphology and the endodontic procedures (for root canal debridement) with the aim being to avoid removing too much tooth structure. Increasing the diameter of the post does not provide a significant increase in the retention of the post; it only increases the stiffness of the post at the expense of the remaining dentine and the fracture resistance of the root. There is little indication for increasing the post width beyond the endodontic cleaning and shaping procedures for all post systems. The post diameter needs to be controlled to preserve the radicular dentine, reduce the potential for perforations, and permit the tooth to resist fracture. However, endodontic treatment often involves wide flared canals where cast posts are the standard method of treatment. These thin-walled canals are then further compromised by the placement of a tapered cast post--thus increasing its risk for fracturing. The Luminex light, transmitting posts provide an alternative treatment option by strengthening weakened, endodontically treated teeth by the combined bonding action of dentine bonding agents and composite resin restorative material. The paper reports on a case where a wide flared root canal was rehabilitated with composite resin using a light-transmitting post. PMID- 15042883 TI - Substance abuse--concerns in dentistry: an overview. PMID- 15042887 TI - General practitioner's radiology case 17. Infected dentigerous cyst. PMID- 15042888 TI - [Three children with general malaise, fever, weight loss and cervical lymphadenopathy]. AB - Combinations of symptoms such as general malaise, fever, weight loss and cervical lymphadenopathy have extensive differential diagnoses. In three children, girls aged 11, 13 and 17 years who presented with these symptoms, three different diagnoses were obtained. The first had Hodgkin's disease, the second mixed connective tissue disease (MCTD), and the third Hodgkin's disease in combination with systemic lupus erythematosus (SLE). A systematic approach is necessary for the diagnosis of such conditions. Careful history taking can provide valuable information while a physical examination provides essential clues for the final diagnosis. In particular, nail-fold lesions, tendon nodules and signs of myopathy should be looked for in patients suspected of MCTD and/or SLE. In Hodgkin's disease, generalized or localised lymphadenopathy combined with a short history of extreme fatigue are the most important. Additional investigations should be individualized in order to minimise the diagnostic delay and make possible early treatment. PMID- 15042889 TI - [From fowl plague to influenza pandemic; a reason for taking precaution]. AB - Throughout Eastern Asia, there is currently an epidemic of fowl plague or highly pathogenic avian influenza, on an unprecedented scale. The prospects for rapid containment are poor. The causative virus, influenza A of the H5N1 subtype, is of limited infectivity for humans. If infection occurs, however, then the consequences are serious and even fatal in a majority of cases. In view of the receptor specificity of avian influenza viruses, this may be related to individually increased susceptibility, which does not lead to further spread. However, it is known that influenza A viruses can readily adapt to replication in the human host by the acquisition of specific gene segments or even by mutations of the avian virus. The extreme scale of human contact with influenza virus of the H5N1 subtype at present engenders fear that there is a high risk of such adaptation and a subsequent pandemic spread. Adequate precautions are necessary, not only in terms of an acceleration of vaccine production but primarily in arranging for sufficient availability of the new antiviral drugs. PMID- 15042890 TI - [Routine funduscopic examination in hypertensive patients not useful]. AB - Funduscopic examination according to the classification of Keith, Wagener and Barker has been used since 1939 to assess retinal damage in patients with hypertension. Improved treatment possibilities and the screening of patients with hypertension seem to have led to a decrease in the incidence and severity of the retinal abnormalities found. The following findings have been published during the past ten years. The assessment of retinopathy is subject to a large interobserver-variability, especially with respect to venous-crossings and arteriolar narrowing of retinal vessels. The positive and negative predictive value of retinopathy for high blood pressure is low. The association of retinopathy with other predictive indicators for organ damage is inconsistent and its association with cardiovascular complications is weak. These findings indicate that the usefulness of funduscopic examination as an instrument for assessing organ damage in patients with hypertension is limited. Therefore it should not be used as a routine diagnostic tool unless a hypertensive crisis is indicated. PMID- 15042891 TI - [Evaluation of frozen sections and cytological diagnosis during surgery]. AB - When a surgeon feels it to be necessary to carry out morphological diagnosis during the operation, the pathologist may assist him or her with rapid cell or tissue diagnosis. This must be a conscious decision to introduce an additional step into the chain of laboratory procedures from the taking of the sample to the reporting of the result. Because this additional phase requires a longer stay in the operating room and additional effort on the part of the laboratory staff, together with an increased risk of misclassification of the disease process, this step should only be taken after careful consideration of the pros en cons. On the one hand, the need for intra-operative diagnosis has decreased because of the availability of better and more accurate pre-operative diagnostic techniques; on the other hand, there are new developments such as the introduction of sentinel node biopsy, in which one must weigh the advantages of a rapid intra-operative diagnosis against the slower but surer results of a more comprehensive lymph-node investigation. Routine examination of the cut edges during an operation is hardly ever done any longer as a result of changed biological knowledge regarding the margins needed for radical surgery. PMID- 15042892 TI - [Physical diagnosis--the Glasgow coma scale for the measurement of disturbances of consciousness]. AB - The Glasgow coma scale (GCS) may be used for the evaluation of disturbances of consciousness. The GCS is a reliable and reproducible test that can easily be carried out at the patient's bedside. The GCS can be used to assess the severity of a head injury and to determine the prognosis of the patient. The prerequisites for a reliable determination of the GCS-score are training of inexperienced personnel, re-assessment by a physician with experience in neurology, correction for external factors and reporting in accordance with a protocol. PMID- 15042893 TI - [From gene to disease: cystinosis]. AB - Cystinosis is an autosomal recessive disorder caused by an impaired transport of cystine out of lysosomes. The most severe infantile form of cystinosis starts with Fanconi syndrome at the age of 3-6 months. Untreated patients develop renal failure before the age of 10. The cystinosis gene (CTNS) maps to chromosome 17p13, spans 23 kb and is composed of 12 exons. CTNS encodes a 367 amino acid protein, cystinosin, which is a H(+)-driven lysosomal cystine transporter. It is enigmatic how lysosomal cystine accumulation induces the clinical symptoms. ATP depletion was demonstrated in an experimental model consisting of loading lysosomes with cystine dimethylester. The amino-thiol cysteamine depletes lysosomal cystine content by a disulfide-exchange reaction with cystine. Therapy with cysteamine should be administered as early as possible and continued after a renal transplantation as the extra renal damage still progresses. Improved life expectancy of cystinotic patients requires the attention of internists with a special interest for this rare disorder. PMID- 15042894 TI - [Diagnostic image (179). A woman with abdominal pain and weight loss. Intestinal infection with Strongyloides stercoralis]. AB - A 40-year-old woman presented with prolonged abdominal complaints, weight loss and iron deficiency anaemia. Coloscopic biopsy revealed a nematode infestation, probably by Strongyloides stercoralis. PMID- 15042895 TI - [The role of cost analysis in the evaluation of the development of medical technology. The case of allogenic stem-cell transplantation]. AB - OBJECTIVE: To estimate the real costs of allogeneic haematopoetic stem-cell transplantation and to compare these with the historically determined budgets that are made available for this purpose ([symbol: see text] 70,038 for genetically related donors and [symbol: see text] 76,826 for unrelated donors). DESIGN: Cost analysis. METHODS: In the period 1994-1999, the direct medical costs (price level of 1998) of bone-marrow transplantation from related donors (BMT), stem-cell transplantation from unrelated donors (VUD-SCT) and allogeneic peripheral-blood stem-cell transplantation (PBSCT) from related donors were determined on the basis of data on adult patients with either acute myeloid leukaemia (n = 66) or acute lymphocytic leukaemia (n = 31). First, the medical resource use by these patients was determined and multiplied by the unit costs of each of the items. Second, a structural programme for allogeneic stem-cell transplantation brings along costs that are not evident from the registration of the medical resource use (e.g., the costs of pretransplantation screening and the selection of the donor). The costs of these items were calculated by taking inventory in the hospitals and assessed by experts. RESULTS: The average costs per transplanted patient were [symbol: see text] 98,334 (BMT), [symbol: see text] 151,754 (VUD-SCT) and [symbol: see text] 98,977 (PBSCT) during the first two years after transplantation. The greater part of the costs was incurred in the transplantation phase. In VUD-SCT, one-third of the total cost was due to the costs of finding a suitable donor. CONCLUSION: The current budget for allogeneic stem-cell transplantation is insufficient to perform the transplantations adequately. Periodic evaluation of the budgets for complicated procedures based on cost analyses has added value for the evaluation of the development of these procedures in time and can thereby contribute to the quality and continuity of care. PMID- 15042896 TI - [A baby boy with cryptorchism, inguinal hernia and internal female genitalia: the persistent Mullerian duct syndrome]. AB - During operation for a right inguinal hernia, a baby boy aged 3 months was discovered to have internal female genitalia. Biopsies were taken from the gonads and blood was sent for karyotyping. The biopsies showed normal testicular tissue and the karyotyping result was 46XY, so the diagnosis of persistent Mullerian duct syndrome (PMDS) was made. Recovery was uneventful. At the age of 2 he underwent bilateral orchidopexy. PMDS is a rare disorder of the MIF synthesis or receptor. Patients present with cryptorchism, inguinal herniation of Mullerian structures, or problems related to the abnormal urinary tract, such as infection or stone formation. In cryptorchism, orchidopexy and life-long palpatory follow up are advised. If urological symptoms occur, surgical removal of the Mullerian remnants may be considered. The prognosis for fertility is poor. PMID- 15042897 TI - [Paraneoplastic syndromes in three patients with renal cell carcinoma]. AB - Renal cell carcinoma was diagnosed in three male patients, 45, 53 and 52 years of age. In addition, they had paraneoplastic symptoms: hypercalcaemia, hyperglycaemia and elevated hepatic enzyme levels, respectively. All three patients underwent tumour nephrectomy, after which the paraneoplastic symptoms disappeared. The first patient died 16 months postoperatively, while the other two were alive and free of symptoms after a follow-up of nine months and four years, respectively. Many patients with renal cell carcinoma remain asymptomatic for a long period of time and 30% of all patients have metastatic disease at the time of diagnosis. The classic triad of flank pain, haematuria and an abdominal mass occurs in only 10% of all cases. However, 20-40% of all patients present with signs of a paraneoplastic syndrome, of which anaemia (20-40%), fever (30%), hypertension (24%), hypercalcaemia (10-15%) and hepatic dysfunction (3-6%) are the most common. PMID- 15042898 TI - [Genetic screening of patients with familial hypercholesterolemia and insurability for life insurance policies and disability cover policies]. AB - In the Netherlands, people with familial hypercholesterolaemia (FH) have been actively screened since 1994 by means of DNA analysis. Recently, the Stichting Opsporing Erfelijke Hypercholesterolemie (Foundation for the Detection of Familial Hypercholesterolaemia) initiated a large scale-screening programme aimed at finding all 40,000 people. The Dutch ministry of Health, Welfare and Sport is providing the financial support. Genetic screening has social implications and raises questions on insurability. The Dutch Medical Examination Act prohibits insurers from posing questions about untreatable, serious inheritable conditions for insured sums under a certain value: for life-insurance policies < [symbol: see text] 150,000 and for disability-cover policies < [symbol: see text] 30,000 in the 1st year and < [symbol: see text] 20,000 in the 2nd year and following years. The Health Council of the Netherlands has defined FH as a serious disease, but one which responds well to treatment. Therefore insurers can request information for the purpose of an accurate risk classification. Insurance contracts can be accepted at normal rates if the target value of LDL-cholesterol < 4 mmol/l and additional risk factors such as smoking and an abnormal BMI are absent; the risk is determined by the phenotype and clinical factors and not by the genotype. PMID- 15042899 TI - [Breast feeding during the first few days after birth: sometimes insufficient]. PMID- 15042900 TI - [Breast feeding during the first few days after birth: sometimes insufficient]. PMID- 15042901 TI - [Breast feeding during the first few days after birth: sometimes insufficient]. PMID- 15042902 TI - [The treatment of ductal carcinoma in situ (DCIS) of the breast]. PMID- 15042903 TI - [The treatment of ductal carcinoma in situ (DCIS) of the breast]. PMID- 15042904 TI - [The treatment of ductal carcinoma in situ (DCIS) of the breast]. PMID- 15042905 TI - [Simplified diagnosis of deep venous thrombosis by applying clinical score and D dimer concentration]. PMID- 15042906 TI - Allergen immunotherapy in historical perspective. PMID- 15042907 TI - Definition of an allergen (immunobiology). PMID- 15042908 TI - Allergen nomenclature. PMID- 15042909 TI - Mechanisms of IgE-mediated allergic reactions. PMID- 15042910 TI - Immunological responses to allergen immunotherapy. PMID- 15042911 TI - Primary and secondary prevention of allergy and asthma by allergen therapeutic vaccines. PMID- 15042912 TI - In vitro tests to monitor efficacy of immunotherapy. PMID- 15042913 TI - Aerobiology. PMID- 15042914 TI - Pharmacoeconomic considerations for allergen immunotherapy. PMID- 15042915 TI - Tree pollen allergens. PMID- 15042916 TI - Grass pollen allergens. PMID- 15042917 TI - Weed pollen allergens. PMID- 15042918 TI - Fungal allergens. PMID- 15042919 TI - Mite allergens. PMID- 15042920 TI - Cockroach and other inhalant insect allergens. PMID- 15042921 TI - Mammalian allergens. PMID- 15042922 TI - Food allergens. PMID- 15042923 TI - Hymenoptera allergens. PMID- 15042924 TI - Biting-insect allergens. PMID- 15042925 TI - Latex allergens. PMID- 15042926 TI - Drug allergens, haptens, and anaphylatoxins. PMID- 15042927 TI - Standardized allergen extracts in the United States. PMID- 15042928 TI - Manufacturing and standardizing allergen extracts in Europe. PMID- 15042929 TI - Preparing and mixing allergen vaccines. PMID- 15042930 TI - Administration of allergen vaccines. PMID- 15042931 TI - Immunotherapy for allergic rhinoconjunctivitis. PMID- 15042932 TI - Allergen immunotherapy: therapeutic vaccines for asthma. PMID- 15042933 TI - Immunotherapy for the prevention of allergic diseases. PMID- 15042934 TI - Immunotherapy for hymenoptera venom and biting insect hypersensitivity. PMID- 15042935 TI - Immunotherapy combined with pharmacotherapy. PMID- 15042936 TI - Immunotherapy in young children. PMID- 15042937 TI - Drug allergy: desensitization and treatment of reactions to antibiotics and aspirin. PMID- 15042938 TI - Non-injection routes for immunotherapy of allergic diseases. PMID- 15042939 TI - Anti-IgE therapy. PMID- 15042940 TI - Modifying allergens and using adjuvants for specific immunotherapy. PMID- 15042941 TI - Novel approaches to immunotherapy for food allergy. PMID- 15042942 TI - Tolerance induced by allergen immunotherapy. PMID- 15042943 TI - Unproven and controversial forms of immunotherapy. PMID- 15042944 TI - Adverse effects and fatalities associated with subcutaneous allergen immunotherapy. PMID- 15042945 TI - Prevention and treatment of anaphylaxis. PMID- 15042946 TI - Instructions and consent forms for allergen immunotherapy. PMID- 15042947 TI - [Should all surgeons perform laparoscopic surgery?]. PMID- 15042948 TI - [Therapeutic endoscopy]. PMID- 15042949 TI - [Laparoscopy in acute abdomen]. PMID- 15042950 TI - [Minimally invasive parathyroidectomy]. PMID- 15042951 TI - [Laparoscopic and endoscopic treatment of reflux]. PMID- 15042952 TI - [Laparoscopic ventral hernia surgery]. PMID- 15042953 TI - [Obesity surgery]. PMID- 15042954 TI - [Laparoscopic treatment of achalasia]. PMID- 15042955 TI - [Endovascular treatment of peripheral arterial obstruction]. PMID- 15042956 TI - [Endovascular treatment of venous occlusions]. PMID- 15042957 TI - [Laparoscopic vascular surgery]. PMID- 15042958 TI - [Endovascular treatment of patients with carotid stenosis]. PMID- 15042959 TI - [Laparoscopic treatment of ectopic pregnancy]. PMID- 15042960 TI - [Minimally invasive surgery in the treatment of cervix cancer]. PMID- 15042961 TI - [Surgical treatment of stress incontinence]. PMID- 15042962 TI - [Endoscopic surgery in lung cancer]. PMID- 15042963 TI - [Minimally invasive bypass surgery without heart-lung-machine]. PMID- 15042964 TI - [Functional endoscopic sinus surgery]. PMID- 15042965 TI - [Minimally invasive endoscopic surgery in the treatment of laryngeal cancer]. PMID- 15042966 TI - [Minimally invasive surgery in the treatment of brain tumors]. PMID- 15042967 TI - [Endovascular treatment of intracranial diseases]. PMID- 15042968 TI - [A million to support handwashing]. PMID- 15042969 TI - [Wasted research continuity and lost hours!]. PMID- 15042970 TI - Update on insulin pump therapy. PMID- 15042971 TI - Treatment of avulsed teeth by school nurses. PMID- 15042972 TI - Pertussis (whooping cough). A genie released from the lamp. PMID- 15042973 TI - Stop everything. Your help is needed. Stay calm. Someone is having a seizure.... PMID- 15042974 TI - Asthma screening. A research utilization project. PMID- 15042975 TI - Obesity: possible solutions. PMID- 15042976 TI - Nursing assessment and treatment of headaches in children. PMID- 15042978 TI - Evaluation & management of trigeminal nerve injuries. PMID- 15042977 TI - New York Society of Forensic Dentistry. PMID- 15042979 TI - Novel risk factors for coronary heart disease. PMID- 15042980 TI - The challenge of restoring the edentulous arch. PMID- 15042981 TI - All ceramic crowns & teeth hard lessons from hard materials. PMID- 15042982 TI - [Rachel Bocher, "toward a retreat to plan a hospital in 2007"]. PMID- 15042983 TI - [Cross infections in advance. But what is being done to prevent them?]. PMID- 15042984 TI - [Allergies and hypersensitivities in children and adults]. PMID- 15042985 TI - [Who takes charge of allergies?]. PMID- 15042986 TI - [Do you recognize chronic obstructive lung disease (COPD)?]. PMID- 15042987 TI - [Type 2 diabetes: blood glucose self-monitoring and observance of dietetic and therapeutic hygiene]. PMID- 15042989 TI - [Hospitalization at home: "The announced orientations must be rapidly concrete"]. PMID- 15042988 TI - [Analyses of medical biology]. PMID- 15042990 TI - [Hospitalization of children, reasons for anger]. PMID- 15042991 TI - [Accept the inevitable. "Put the separation in concrete form"]. PMID- 15042992 TI - [Imatinib. An anticancer agents indicated in the treatment of chronic myeloid leukemia]]. PMID- 15042993 TI - [Nursing care and liberal exercise]. PMID- 15042994 TI - [Difficulties told by a nurse at the end of the life of an infant during resuscitation]. PMID- 15042995 TI - Allele frequencies of four STR loci in Eastern Chinese population. PMID- 15042996 TI - Northwest Italian population data for thirteen tetrameric and two pentameric STR loci. PMID- 15042997 TI - Distributions of allelic frequencies and haplotypes of two new STR loci in a Chinese Han population. PMID- 15042998 TI - Population genetics of two STR loci D2S1396 and D6S474 in a Chinese population. PMID- 15042999 TI - Molecular epidemiology of cystic echinococcosis. AB - Echinococcus granulosus exhibits substantial genetic diversity that has important implications for the design and development of vaccines, diagnostic reagents and drugs effective against this parasite. DNA approaches that have been used for accurate identification of these genetic variants are presented here as is a description of their application in molecular epidemiological surveys of cystic echinococcosis in different geographical settings and host assemblages. The recent publication of the complete sequences of the mitochondrial (mt) genomes of the horse and sheep strains of E. granulosus and of E. multilocularis, and the availability of mt DNA sequences for a number of other E. granulosus genotypes, has provided additional genetic information that can be used for more in depth strain characterization and taxonomic studies of these parasites. This very rich sequence information has provided a solid molecular basis, along with a range of different biological, epidemiological, biochemical and other molecular-genetic criteria, for revising the taxonomy of the genus Echinococcus. This has been a controversial issue for some time. Furthermore, the accumulating genetic data may allow insight to several other unresolved questions such as confirming the occurrence and precise nature of the E. granulosus G9 genotype and its reservoir in Poland, whether it is present elsewhere, why the camel strain (G6 genotype) appears to affect humans in certain geographical areas but not others, more precise delineation of the host and geographic ranges of the genotypes characterised to date, and whether additional genotypes of E. granulosus remain to be identified. PMID- 15043000 TI - The effect of forefoot varus on postural stability. AB - STUDY DESIGN: Counterbalanced experimental design study comparing a group of subjects with greater than or equal to 70 of forefoot varus (MFV) to a group with less than 70 of forefoot varus (LFV). OBJECTIVES: To investigate the effect of forefoot varus on single-limb stance postural stability (PS). BACKGROUND: Impaired PS has been implicated as a potential risk factor for sustaining acute foot and ankle injuries. The identification of variables that deleteriously affect PS may be important in the prevention of future injuries. METHODS AND MEASURES: Postural stability of the MFV group (n = 20) and the LFV group (n = 12) was assessed during right and left single-limb stance and eyes-open and eyes closed conditions. Standard deviations of the x-axis and y-axis ground reaction forces measured via a force platform were used to represent anteroposterior (AP) and mediolateral (ML) PS, respectively. The mean of 3 successful 5-second trials of each testing condition was calculated and used for subsequent data analysis using 3-way mixed-model ANOVAs with 1 between-subject and 2 within-subject factors. RESULTS: The AP PS scores of the MFV group were significantly greater than those of the LFV group (P < .05). ML PS scores, although higher in the MFV group, were not significantly different from those of the LFV group. Both groups had significantly greater AP and ML PS scores during the eyes-closed versus the eyes-open condition (P < .05). CONCLUSIONS: The results suggest that the presence of greater than or equal to 70 of forefoot varus may significantly impair AP PS. The decreased stability associated with increased forefoot varus may be due to decreased joint congruity and consequently an increased reliance on soft tissue structures for stability. PMID- 15043001 TI - Detection and isolation of H1N1 influenza virus from pigs in Korea. PMID- 15043002 TI - Rapid gas chromatography-mass spectrometry screening method for human pharmaceuticals, hormones, antioxidants and plasticizers in water. AB - A rapid gas chromatography-mass spectrometry (GC-MS) method was developed and validated allowing quantification at the ng/l level of 19 analytes in water including human pharmaceuticals, hormones, antioxidants and a plasticizer. On line continuous liquid-liquid extraction with dichloromethane of 10-401 unfiltered water samples was used to achieve a 10000-40000-fold concentration factor. No sample cleanup or derivatization was required. Recoveries ranged from 57 to 120%. Application of the method to water recycling plant effluent demonstrated the presence of nearly all targeted compounds at ng/l to microgram/l levels. Screening for nontarget compounds in the treated effluent samples indicated the method could be readily extended to include additional analytes. PMID- 15043003 TI - Nonpharmacological suppression of oestrus in the mare. PMID- 15043004 TI - Coverage and costs of mass immunization of an oral cholera vaccine in Vietnam. AB - The objective of this study was to describe a mass-immunization campaign of a locally-produced oral, killed whole-cell cholera vaccine in Hue city, Vietnam. Mass immunization with a 2-dose regimen of the vaccine was conducted in 13 communes in early 1998. The total, age- and sex-specific vaccine coverage was calculated using data from the vaccination records and the government census. The number of vaccine doses procured, administered, wasted, and left over, and the human and other resources required to prepare and conduct the vaccination campaign were systematically recorded. Government expenditure for planning, procurement, and delivery of the vaccine were documented. In total, 118,555 (79%) of the 49,557 targeted population were fully vaccinated during the mass vaccination campaign. The total expenditure for the project was US dollar 105,447, resulting in a cost per fully-vaccinated person of US dollar 0.89. Mass immunization with this locally-produced oral, killed cholera vaccine was found to be feasible and affordable with attainment of high vaccination coverage. PMID- 15043005 TI - The physiology of channel swimmers. 1955. PMID- 15043006 TI - Oxygen therapeutics: can we tame haemoglobin? AB - Chemically modified or genetically engineered haemoglobins (Hbs) developed as oxygen therapeutics (often termed 'blood substitutes') are designed to correct oxygen deficit due to ischaemia in a variety of clinical settings. These modifications are intended to stabilize Hb outside its natural environment--red blood cells--in a functional tetrameric and/or polymeric form. Uncontrolled haem mediated oxidative reactions of cell-free Hb and its reactions with various oxidant/antioxidant and cell signalling systems have emerged as an important pathway of toxicity. Current protective strategies designed to produce safe Hb based products are focused on controlling or suppressing the 'radical' nature of Hb while retaining its oxygen-carrying function. PMID- 15043007 TI - Virus entry: molecular mechanisms and biomedical applications. AB - Viruses have evolved to enter cells from all three domains of life--Bacteria, Archaea and Eukaryotes. Of more than 3,600 known viruses, hundreds can infect human cells and most of those are associated with disease. To gain access to the cell interior, animal viruses attach to host-cell receptors. Advances in our understanding of how viral entry proteins interact with their host-cell receptors and undergo conformational changes that lead to entry offer unprecedented opportunities for the development of novel therapeutics and vaccines. PMID- 15043008 TI - Effectiveness of prostaglandin E1 in relieving obstruction in coarctation of the aorta without opening the ductus arteriosus. AB - The two main theories regarding the pathogenesis of coarctation of the aorta are the Skodaic hypothesis of ductal tissue constricting the aorta at the level of the insertion of the ductus arteriosus and the flow theory of decreased ascending aortic blood flow in the fetus, which results in associated isthmic narrowing and a localized shelf. To document that ectopic ductal tissue constriction can cause coarctation of the aorta in the absence of a patent ductus arteriosus, we report three cases of infants presenting with critical coarctation who responded to prostaglandin E1 infusion without opening the ductus arteriosus. PMID- 15043009 TI - Making state public health laws work for SARS outbreaks. PMID- 15043010 TI - Medicare overhaul of 2003. PMID- 15043011 TI - Steps in planning for change. PMID- 15043012 TI - Focusing on quality of care. PMID- 15043013 TI - Use of an Amplatzer duct occluder for closing an aortico-left ventricular tunnel in a case of noncompaction of the left ventricle. AB - Aortico-left ventricular tunnel (ALVT) is a very rare congenital anomaly. Noncompaction of the left ventricle or "spongy myocardium" is another rare congenital cardiomyopathy. We report probably the first case in which these two rare anomalies causing pump failure coexisted in a 4-year-old make and the tunnel was successfully treated nonsurgically. Transcatheter closure of ALVT with the Amplatzer duct occluder is feasible and safe in a case of noncompaction of the LV with reduced LV function. PMID- 15043014 TI - Protein-losing enteropathy: radiofrequency fenestration of the atrial septum after failure of transseptal needle puncture. PMID- 15043015 TI - SARS epidemic in the press. PMID- 15043016 TI - SARS-associated coronavirus infection in teenagers. PMID- 15043017 TI - SARS preparedness and response planning. PMID- 15043018 TI - HIV on the increase in UK. PMID- 15043019 TI - Are there genes for aneurysm in the blueprint of the human genome? PMID- 15043020 TI - Peroneal neuropathy due to a popliteal aneurysm in a patient with infectious endocarditis. AB - We present an 18-year-old young man with the diagnosis of infective endocarditis who also suffered from concomitant knee pain. The clinical examination revealed right homonymous hemianopsia and left peroneal neuropathy. Magnetic resonance imaging studies of the knee demonstrated a lobulated saccular aneurysm in the left popliteal fossa. We diagnosed this patient as having peripheral mononeuropathy due to an aneurysm compressing the peroneal nerve in the left poplitea. PMID- 15043021 TI - Behcet's disease: treatment of popliteal pseudoaneurysm by an endovascular stent graft implantation. AB - Behcet's disease is a multisystemic chronic autoimmune process that can be responsible for some vascular lesions, in addition to its typical dermatologic lesions. Arterial pseudoaneurysm formation can be diagnosed in this group of patients and generally surgery is indicated. In this report, we discuss an alternative endovascular intervention and its advantages. An endovascular stent graft was placed in the popliteal artery pseudoaneurysm of a 37-year-old male with Behcet's disease. Early results of this procedure are good thus far. Implantation of an endovascular stent graft, an alternative modality in the treatment of pseudoaneurysms, can result in a shorter hospital stay and a significantly shorter period before returning to normal life. General anesthesia and surgical dissection are avoided, thus diminishing morbidity. PMID- 15043022 TI - Popliteal artery pseudoaneurysm caused by osteochondroma. AB - A 49-year-old man with asymptomatic osteochondroma was found to have a pseudoaneurysm of the left popliteal artery. An angiography showed a popliteal artery pseudoaneurysm adjacent to a femoral osteochondroma. The osteochondroma was excised and the popliteal artery containing the tear was also excised by end to-end anastomosis. Vascular complication of osteochondroma is extremely rare. Paul reported the first case of a popliteal artery pseudoaneurysm due to osteochondroma in 1953. A computerized literature search revealed 40 additional cases reported in English, providing sufficient detail to allow comparison. The average age of the patients was 22.6 years (range 9-51) and most cases identified involved males (30 of 41, 73%). Most pseudoaneurysm were located in the popliteal artery. This report discusses the authors' experience with this disorder and a review of the literature in English. PMID- 15043024 TI - Deep venous thrombosis caused by congenital absence of inferior vena cava, combined with hyperhomocysteinemia. AB - We present a case of suprarenal and infrarenal absence of the inferior vena cava, combined with hyperhomocysteinemia in a 39-year-old woman who presented with symptoms of deep venous thrombosis. The patient also had a homozygous mutation of C677T methylenetetrahydrofolate reductase. Deep vein thrombosis has a multifactorial etiology involving both genetic and acquired factors. Absence of the inferior vena cava is a rare congenital anomaly, but recently it was confirmed as an important risk factor for the development of deep vein thrombosis, especially in young persons. Hypercoagulability due to hyperhomocysteinemia with a tendency toward venous stasis, mediated by congenital absence of the inferior vena cava is thought to have caused deep vein thrombosis in our patient. To our knowledge, this association has not yet been reported. The clinical features and prognosis of the entity are discussed. PMID- 15043025 TI - Mycotic aneurysms affecting both lower legs of a patient with Candida endocarditis--endovascular therapy and open vascular surgery. AB - The purpose of this study was to report the endovascular and open surgery treatment of Candida-associated mycotic aneurysms in both lower limbs. A 53-year old patient suffering from Candida endocarditis following aortic valve replacement developed mycotic aneurysms in both lower limbs. The angiography revealed a large aneurysm of the tibioperoneal trunk affecting the right leg. In the left leg, sacculation had developed in section III of the popliteal artery. The right aneurysm was obliterated by embolization with coils. On the left side, the large aneurysm of the popliteal artery was resected; vascular continuity was re-established by interposing a segment of the patient's greater saphenous vein. The postoperative course was uneventful. Mycotic aneurysm is a rare disease. A site in the crural vessels is regarded as exceptionally seldom. To our knowledge, no Candida-associated mycotic aneurysm has been described in this region before. Both endovascular treatment and open surgery proved to be successful. PMID- 15043023 TI - [Therapy of malignant melanoma at the stage of distant metastasis]. AB - Treatment of melanoma in the stage of distant metastasis aims on palliation and achievement of durable tumor remission with prolongation of survival. As long as metastasis is confined to one organ system and is removable, surgery remains the treatment of first choice. In limited metastasis radiotherapy may likewise be indicated, particularly in bone and brain metastasis. More extensive metastasis should be treated by chemotherapy or chemoimmunotherapy. Monochemotherapy with dacarbazine, temozolomide, fotemustine and vindesine or its combinations with interferon-alpha are currently preferred. Polychemotherapy or its combinations with interferon-alpha and interleukin-2 are suitable to produce higher response rates but failed to prolong survival. As these treatments are associated with substantially higher toxicity they have been widely abandoned. Combined treatment with dacarbazine and interferon-alpha obtain tumor responses or stable disease in 40-50% and objective tumor remissions in 15-20% of patients. Effective cancer vaccination strategies and blockade of melanoma specific target molecules are currently developed as new treatment options. PMID- 15043026 TI - [Undesirable effects of corticosteroids]. PMID- 15043027 TI - [Specification of therapy is the key point for raising the effect for amblyopia]. PMID- 15043028 TI - [Method and evaluation for calculating logarithm of visual angle]. PMID- 15043030 TI - Microaerophilic bacteria transduce energy via oxidative metabolic gearing. AB - Heterotrophic microaerophilic bacteria, a diverse and ubiquitous group, are specialized for growth in O2 limited environments. Until recently, microaerophilic bacteria have been grouped with obligate aerobic organisms whose metabolic rates slow in response to physiological O2 limitation. In contrast, microaerophilic bacteria are adapted to maintain essentially constant turnover of primary energy substrates in response to a wide range of physiological O2. This capacity, oxidative metabolic gearing, allows microaerophilic bacteria to maintain catabolic enzymes, substrates, and cofactors at high steady-state levels. Oxidative metabolic gearing is thus adaptive, as it allows microaerophilic bacteria to respond to changes in physiological O2 relatively rapidly when compared to aerobic, anaerobic, or facultative microorganisms. PMID- 15043031 TI - Vector-omics? PMID- 15043032 TI - [The 92nd annual meeting of the Japanese Urological Association. Osaka, Japan. April 10-12, 2004. Abstracts]. PMID- 15043033 TI - Ovarian cancer. PMID- 15043034 TI - Using the BETTER Model to assess sexuality. PMID- 15043036 TI - The management of asthma in the UK must be reassessed. PMID- 15043035 TI - Shift pattern problems make nurses leave NHS. PMID- 15043037 TI - Bioelectric power generation. AB - In the recent article by Chaudhuri and Lovley, a fuel cell is described in which a microorganism, Rhodoferax ferrireducens, is used to oxidize glucose to carbon dioxide at neutral pH. This reaction occurs via direct bioelectrocatalysis: the microorganism uses the anode itself as the terminal oxidant to which liberated electrons are transferred, and does so with 83% efficiency. These findings are significant because they demonstrate a new approach for harvesting energy from the environment using microorganisms. PMID- 15043038 TI - Report calls for more postmortems as critical diagnoses are being missed. PMID- 15043039 TI - People with learning disabilities are valued. PMID- 15043040 TI - People with learning disabilities are valued. PMID- 15043041 TI - A new solution for improving gene delivery. AB - Intense research has focused on methods for overcoming the various barriers to DNA delivery while simultaneously reducing toxicity. Recently, Yuan and colleagues discovered a 'leakage route', whereby viral vectors escaped the injection site and entered the systemic circulation and contributed to the systemic toxicity in cancer gene therapy. They also demonstrated that alginate solution sealed this leak. Although their work focused on intratumoral injection of viral vectors, the findings can be extended to drug delivery in general, particularly DNA delivery. PMID- 15043042 TI - Leadership can be found within nursing. PMID- 15043043 TI - The cell equator - more than poles apart. AB - Cytokinesis is the last ritual of a dividing cell. Determining the position and horizon of the cell furrow is important for equal distribution of cytoplasmic content between the two daughter cells. The traditional view promotes a classical sequence of bipolar spindle formation followed by cytokinesis. However, a new understanding has recently emerged that uncouples these events. The cell-furrow formation seems to ignore spindle polarity and is instead dependent on the stability and dynamics of cortical microtubules. PMID- 15043044 TI - Engineered plants with elevated vitamin E: a nutraceutical success story. AB - Vitamin E has been touted as a panacea for age-related diseases, including cardiovascular disease and Alzheimer's disease and, thus, the demand for this nutraceutical has increased dramatically in recent years. This demand has, in turn, driven research to increase vitamin E production from plant sources. We have summarized the cumulative work of several groups in this area, describing the current status of efforts to bioengineer plants for elevated vitamin E content. PMID- 15043045 TI - How can genetically modified foods be made publicly acceptable? AB - A recent study by Lusk suggests that consumers might voluntarily pay more for a genetically modified (GM) food than a non-GM equivalent if made aware of the possible health benefits. However, other research indicates that the acceptability of novel hazards is affected by a variety of factors, in addition to benefits, and that making agricultural biotechnology publicly acceptable will be more complex than indicated by the results from Lusk's study. PMID- 15043046 TI - Introduction science in early modern east Asia: State patronage, circulation,and the production of books. PMID- 15043047 TI - The Golden Mirror in the imperial court of the Qianlong emperor, 1739-1742. AB - In the last month of 1739, the third f the Manchu rulers, the Qianlong emperor (r. 1736-1795), ordered the compilation of a treatise on medicine "to rectify medical knowledge" throughout the empire. By the end of 1742, eighty participants chosen from several offices within the palace bureaucracy based in Beijing completed the Golden Mirror of the Orthodox Lineage of Medicine, the only imperially commissioned medical text the Qing government's Imperial Printing Office published. The Golden Mirror represents both the limitations in the power of the Qianlong emperor and the dominance in the Manchu court of Chinese scholarship from the Jiangnan region during the first decade of his reign. Chinese scholars participating in the compilation of the Golden Mirror fashioned a medical orthodoxy for the empire in the mid-eighteenth century from regional trends since the sixteenth century. The Golden Mirror is an illuminating example of how medical scholars participated in the formation of evidential scholarship in early-modern China and why Manchu patronage, southern Chinese scholarship, and medical orthodoxy, coalesced in the imperial court of the Qianlong emperor. PMID- 15043048 TI - When science develops outside state patronage: Dutch studies in Japan at the turn of the nineteenth century. AB - It is one of the peculiar features of the movement of translation of Western scientific treatises from Dutch into Japanese, known as Dutch learning (rangaku), that if first originated in Nagasaki with a group of Japanese interpreters. This group differed from the scholarly community of the capital, Edo, by both training and social status. This article shows how this difference contributes to explaining some of the particularities of rangaku in its initial phase. A case in point is Shizuki Tadao's introduction of Newtonian physics and astronomy. Yet, Sugita Genpaku, a major representative of the Edo scholarly community, gave an account of the beginning of Dutch learning that attempted to minimise, or even to erase, the contribution of the Nagasaki interpreters, who were dismissed as unscholarly. This attempt, too, is best understood in the light of the difference between the two communities. PMID- 15043049 TI - [Saints as protectors against falling sickness]. AB - In Christian Europe of the High Middle Ages, saints played a central role in the everyday life of the ailing. Alongside healing attempts which involved magic and/or scientifically-based medicine, the invocation of specific patron saints for protection against evils or for the curing of ailments was a widespread practise. A large choice of patron saints was "available" for a wide range of diseases, especially those nowadays classified as neurologic or psychiatric. For the falling sickness alone, e.g., there is evidence of some twenty patron saints reputed to have a particular involvement. Surprisingly, there is no evidence of a comparable devotion to patrons for apoplectics. This "negative result"is confirmed by a thorough examination of medieval sources. St. Wolfgang and St. Andreas Avellino are the only two proven stroke patrons. Both, however, were only known within their respective locations. The absence of a specific supportive Christian figure for stroke victims deserves particular analysis: The high fatality rate of apoplexy and the lack of commercial interest on the part of the Christian places of pilgrimage may serve as possible explanations. PMID- 15043050 TI - Re: Retraction request: AJNR 23: 868-870, May 2002. PMID- 15043052 TI - Current world literature. Nephrology. PMID- 15043051 TI - A new instrument for pain assessment in vulvar vestibulitis syndrome. AB - Vulvar vestibulitis syndrome (VVS) is a common form of dyspareunia in premenopausal women. The standard test for diagnosing VVS is the cotton-swab test, during which a cotton-swab is applied to various locations of the vulvar vestibule. However, there is much variation in the implementation of this test relating to the precise vestibular locations palpated, the order of palpation, and the force used during palpation. We introduce a new simple, mechanical device, a vulvalgesiometer, to standardize genital pain assessment and present promising preliminary data from women with VVS and nonaffected women. These data indicate that women with VVS have significantly lower vestibular pain thresholds compared with control women. During painful vulvar stimulation with the vulvalgesiometer, women with VVS described the pain with adjectives similar to those used to describe their intercourse pain (e.g., burning). This novel device has several important implications for genital pain measurement in women who suffer from urogenital pain. PMID- 15043053 TI - Current world literature. Therapeutics and toxicology. PMID- 15043055 TI - Class III nucleotide cyclases in bacteria and archaebacteria: lineage-specific expansion of adenylyl cyclases and a dearth of guanylyl cyclases. AB - The Class III nucleotide cyclases are found in bacteria, eukaryotes and archaebacteria. Our survey of the bacterial and archaebacterial genome and plasmid sequences identified 193 Class III cyclase genes in only 29 species, of which we predict the majority to be adenylyl cyclases. Interestingly, several putative cyclase genes were found to have non-conserved substrate specifying residues. Ancestors of the eukaryotic C1-C2 domain containing soluble adenylyl cyclases as well as the protist guanylyl cyclases were found in bacteria. Diverse domains were fused to the cyclase domain and phylogenetic analysis indicated that most proteins within a single cluster have similar domain compositions, emphasising the ancient evolutionary origin and versatility of the cyclase domain. PMID- 15043054 TI - Paracelsianism and the orthodox lutheran rejection of vital philosophy in early seventeenth-century Denmark. AB - Paracelsian medicine and natural philosophy was formed during the Radical Reformation and incorporated metaphysical propositions that were incompatible with the Lutheran confession as codified in the Confessio Augustana and elaborated in the ultra-orthodox Formula of Concord. Although Paracelsian ideas and practices were endorsed by important philosophers and physicians in late sixteenth century Denmark without raising serious alarm, the imposition of strict Lutheran orthodoxy in the Danish Church and a concomitant resurgence of Aristotelian philosophy drew attention to the religious heterodoxies inherent in Paracelsianism. Unacceptable theological and religious propositions, which reached Denmark in Rosicrucian texts and were implicit in certain medical and philosophical treatises, were in many cases inseparable from core Paracelsian concepts, with the result that Danish academic philosophers, physicians, and theologians rejected Paracelsian ideas except where they could be accommodated to acceptable Galenic and Aristotelian interpretations. When this was done, such ideas are arguably no longer Paracelsian in any meaningful way. PMID- 15043056 TI - Plague and more plagues. [Review of: Cohn, SK, Jr. The black death transformed: disease and culture in early renaissance Europe. London, Edward Arnold, 2003; Scott S and Duncan CJ. The biology of plagues: evidence from historical populations. London, Cambridge University Press, 2001]. PMID- 15043057 TI - An investigation of genetic counselors' discussion of genetic discrimination with cancer risk patients. AB - Twenty-five genetic counselors who see familial cancer risk patients were interviewed about their definitions of genetic discrimination, perceptions of patient risk for discrimination, frequency and type of discrimination experienced by their patients, sources of information about discrimination, and what they tell patients about genetic discrimination. There was variability in participant definitions and content of discussions of genetic discrimination. Although 82% regarded risk to be low to theoretical, 40% reported possible instances of genetic discrimination, including insurance companies requiring results to cover procedures, denial of life/health insurance, social discrimination, and employment discrimination. Ninety-six percent of the sample reported always or almost always discussing genetic discrimination, and one or more of the following: insurance discrimination risks, protective legislation, no known cases positively documenting discrimination, strategies to avoid discrimination, and uncertainty of protective measures. The majority use current literature and research as information sources. Practice, policy, and research implications are discussed. PMID- 15043058 TI - Traditions of pathology in Western Europe. Theories, institutions and their cultural setting: An introduction. PMID- 15043059 TI - Spelling sickness: The aufschreibesystem of medical semiotics in the eighteenth century. PMID- 15043060 TI - The psychological dimension of informed consent: dissonance processes in genetic testing. AB - This paper discusses the issue of the psychological dimension of informed consent. In this paper, the author proposes that informed consent is a continuous variable rather than a dichotomous one. When clients better understand their motives and actual, rather than just perceived degree of choice in pursuing a particular option in a medical setting, their level of informed consent is greater. Findings from existing literature in the field of genetic testing are examined in terms of dissonance theory. These findings suggest that testing candidates sometimes overestimate their coping skills and minimize the threat to psychological integrity that a particular genetic result may pose. Counseling directed towards realistic appraisal of degree of choice in pursuing testing is examined as an aspect of supporting informed consent and possibly reducing the potential for adverse psychological outcome in the longer term. PMID- 15043061 TI - "Realpolitik": Friedrich von Recklinghausen's pathology for the kaiserreich. PMID- 15043062 TI - "A general rule in the post-mortem finding of suicides?" German pathology and forensic medicine and the suicide problem 1900-1945. PMID- 15043063 TI - The representational framework of health and disease. PMID- 15043064 TI - German pathology and the defence of autopsy since 1850. PMID- 15043065 TI - Current world literature. Antimicrobial agents: bacterial/fungal. PMID- 15043066 TI - Current world literature. Antimicrobial agents: parasitic/viral. PMID- 15043067 TI - Current world literature. Infections of the immunocompromised host. PMID- 15043068 TI - Mitochondrial DNA extracted from the British Columbia "Iceman". PMID- 15043069 TI - The Capestrano warrior: Artistic caprice or disease? PMID- 15043070 TI - The African Nutrition Graduate Students Network: a new initiative for nutrition in Africa. PMID- 15043071 TI - Radial versus right internal thoracic artery for myocardial revascularization. PMID- 15043072 TI - Cosmology: synchrotron radiation and quantum gravity. PMID- 15043073 TI - Enamelomas in paleopathology: A report of three cases and a review of the literature. PMID- 15043074 TI - Malaria and porotic hyperostosis in the Aegean world from the Paleolithic to the Geometrical period. PMID- 15043075 TI - Surgical correction of phimosis in the etruscan period. PMID- 15043077 TI - Lung volume reduction surgery for severe emphysema. PMID- 15043076 TI - FDG positron emission tomography for evaluating breast cancer. PMID- 15043078 TI - Magnetic resonance imaging of the breast in screening women considered to be at high genetic risk of breast cancer. PMID- 15043079 TI - Percutaneous intradiscal radiofrequency thermocoagulation for chronic discogenic low back pain. PMID- 15043080 TI - Radiofrequency ablation of unresectable hepatic tumors. PMID- 15043081 TI - Special report: left ventricular assist devices as destination therapy for end stage heart failure--cost-effectiveness analysis. PMID- 15043082 TI - Transesophageal endoscopic treatments for gastroesophageal reflux disease. PMID- 15043083 TI - Use of intermittent or continuous interstitial fluid glucose monitoring in patients with diabetes mellitus. PMID- 15043084 TI - Wireless capsule endoscopy for small-bowel diseases other than obscure GI bleeding. PMID- 15043085 TI - [Research and control of Hantaan virus and HFRS]. PMID- 15043086 TI - The young adult with hip pain: diagnosis and medical treatment, circa 2004. AB - Hip pain in young adults (18-35 years old) often is characterized by nonspecific symptoms, normal imaging studies, and vague findings from the history and physical examination. In younger patients, pain is more likely to be caused by congenital hip dysplasia, athletic injuries, trauma, spondyloarthropathy, and by conditions that first appear during this stage of life, such as rheumatoid arthritis, osteoarthritis, intravenous drug use, alcoholism, or corticosteroid use. The history and physical examination may narrow the diagnosis to intraarticular, extraarticular, or referred sources of pain. Plain radiography and magnetic resonance imaging are the preferred initial imaging procedures. Analyses of the blood, urine, and synovial fluid can be helpful in diagnosing inflammation, infection, and systematic rheumatic disease. Fractures, infection, and ischemic necrosis should be ruled out early because they require immediate treatment to prevent damage to the joint. Hip trauma at a young age increases the risk of osteoarthritis with advancing age, and, unlike most older adults, young adults receiving total hip replacement can expect revision surgery. Medical treatment often involves patient education, physical therapy, and pharmacotherapy. Acetaminophen, nonsteroidal anti-inflammatory drugs, and opioids for pain and antibiotics for infection are the most often prescribed drugs for this population. PMID- 15043087 TI - Clinical and radiographic assessment of the young adult with symptomatic hip dysplasia. AB - Acetabular dysplasia is a major precursor of osteoarthritis of the hip. It is important to identify patients before the development of osteoarthritis, because these patients may be candidates for joint-preserving surgery, such as pelvic osteotomy. Patients with hip dysplasia, and no osteoarthritis, present with unique findings on history and physical examination. On history, patients should be questioned for symptoms unique to dysplasia, such as locking, snapping, and feelings of instability. Physical examination should look for signs of labral disease, by using the impingement test. Patients suspected of having symptomatic dysplasia should have plain radiographs, including an anteroposterior radiograph of the pelvis taken with the patient standing, a false profile view, and an abduction view of the hip. The anteroposterior radiograph of the pelvis and false profile view will help to quantify the degree of the dysplasia. The abduction view is important in determining the patient's eligibility for joint-preserving hip surgery. Computed tomography scanning and magnetic resonance imaging are reserved for patients who are operative candidates. Computed tomography scanning can aid in the planning of complex pelvic osteotomies. Magnetic resonance imaging especially is useful for delineating labral disease. It is this combination of history, physical examination, and radiographic workup that will allow successful treatment of the dysplastic hip before the development of osteoarthritis. PMID- 15043088 TI - Survivorship analysis of surgical treatment of the hip in young patients. AB - Survivorship analysis has become the standard statistical technique for evaluating the results of total joint arthroplasty. There are several problems associated with survivorship analysis as it is presented in the orthopaedic literature that apply to younger and older patients. Studies conducted within the same institution using the same prosthesis have reported differences between young and old patients as high as 30% whereas other studies have reported differences as low as 2%. When discrepancies such as this exist it is clear that more care is needed when presenting results and making comparisons among younger patients. The major reason for the higher revision rates in younger patients usually has been an assumption of higher patient activity resulting in more wear for the prosthesis. However, some older patients are very active after surgery and some younger patients are very inactive. I will discuss the need for more sophisticated instruments measuring patient activity and statistical procedures that are necessary for the proper presentation of data involving younger patients. The statistical issues include the need for multivariate survival analysis and the need for time varying measurements of patient activity. PMID- 15043089 TI - Core decompression for osteonecrosis of the hip. AB - The ultimate goal of treatment of patients with osteonecrosis of the hip is preservation of the femoral head. However, the development of a successful strategy to treat this disease has been difficult because neither the etiology nor the natural history of osteonecrosis of the hip has been defined clearly. Core decompression of the hip is one of the most commonly done surgical procedures to treat the early stages of osteonecrosis of the femoral head. However, there is no general consensus among investigators regarding either the specific indications for this procedure, or the specific technique of core decompression that would optimize results. The data available suggest that core decompression is more effective than nonoperative treatment particularly for patients with limited involvement of the femoral head and with an area of sclerosis surrounding the necrotic bone. The combination of core decompression with the use of osteoinductive or angiogenic agents to enhance bone formation and repair may enhance the results of this surgical procedure in the future. PMID- 15043090 TI - Femoral osteotomies for avascular necrosis of the femoral head. AB - Avascular necrosis of the femoral head affects 10,000 to 20,000 people every year and often these patients are in their 30s and 40s. The natural history of this disease is important to understand because the rate of femoral head collapse and treatment options are related to multiple factors including the cause of the disease, its stage at initial presentation, and the size and location of the lesion. Various treatment options have been purposed for different stages of avascular necrosis of the femoral head. Nonoperative treatments include protected weightbearing, electrical stimulation, and pharmacologic treatments. Operative treatment includes core decompression with or without grafting, debridement and grafting, osteotomy, fusion, hemiresurfacing, hemiarthroplasty, and total hip arthroplasty. The purpose of the current study was to discuss the history, indications, techniques, and results of proximal femoral osteotomies for the treatment of avascular necrosis of the femoral head. PMID- 15043091 TI - Impact of implant design on femoral head hemiresurfacing arthroplasty. AB - Two consecutive series of cemented femoral hemiresurfacing arthroplasty for patients with Ficat Stage III and early Stage IV osteonecrosis of the femoral head were studied to identify the impact of design on the clinical and radiographic results. Group I (30 patients, 33 hips) components had a cobalt chrome shell with a tapered inner dimension, no stem, and sizes in 2-mm increments. Group II (37 patients, 51 hips) components differed by adding a proportional stem, increased spherical coverage, and sizes in 1-mm increments. The average age for the patients in both groups was 40 years. Fifty percent of the patients in Group I were men: in 43% of patients osteonecrosis was associated with steroid use and in 21% of patients it was associated with alcohol use. Thirty-three percent of the patients in Group II were men: in 41% of patients osteonecrosis was associated with steroid use and in 17% of patients it was associated with alcohol use. The average followup is 42 months for Group I and 24 months for Group II. Neither group experienced infections, nerve palsies, dislocations, or loosening. In Group I, two patients died of unrelated causes and five patients had reoperations, two for femoral neck fractures, and three for unsatisfactory pain relief. No patients in Group II had femoral neck fractures but three patients had reoperations for unsatisfactory pain relief. The stemmed component in Group II has resulted in an improvement in component position and elimination of femoral neck fractures in this series. PMID- 15043092 TI - Recognizing acetabular version in the radiographic presentation of hip dysplasia. AB - Attention to the radiographic shadow of the walls of the acetabulum seen on the anteroposterior projection of a well-centered radiograph of the pelvis is important to identify the orientation of the opening of the acetabulum. This is referred to as the acetabular version. The opening of the mouth of the acetabulum may be oriented anteriorly, which is referred to as anteverted; posteriorly, or retroverted; or when the shadow of the lips of the anterior and posterior walls terminate together on the lateral edge of the acetabular rim, neutral version. The radiographs of 153 patients with developmental hip dysplasia presenting with hip pain were reviewed to determine the frequency of retroversion of the acetabulum. In this group of patients, retroversion of the hip socket was surprisingly common, one in three hip sockets, and generally was associated with smaller values of the lateral center edge measurement. The observation is important, because it must be taken into account when planning a corrective osteotomy of the acetabulum so the abnormal horizontal orientation of socket is corrected along with the usual anterior and lateral insufficiency. PMID- 15043093 TI - Abnormal extension of the femoral head epiphysis as a cause of cam impingement. AB - The etiology of an insufficient femoral head-neck offset has not been identified yet. It was investigated whether a decreased head-neck offset might be correlated with an unusual orientation of the physeal capital scar. Therefore, the femoral head-neck offset and the extension of the physeal scar onto the femoral neck were measured with specific magnetic resonance imaging arthrography. The measurements were done in 15 patients with anterior femoroacetabular impingement attributable to a nonspherical head and were compared with 15 age- and gender-matched control subjects. Eight serial magnetic resonance imaging sections perpendicular to the femoral neck axis were used in each hip to measure the head-neck offset and the epiphyseal extension toward the femoral neck at 16 measurement points. In both groups there was an inverse correlation between the amount of head-neck offset and the relative extension of the capital physeal scar in the cranial hemisphere of the head. Within the anterosuperior head quadrant, there was statistically significant different decrease of the head-neck offset and increase of the lateral epiphyseal extension in the patients compared with the control subjects. These findings suggest a growth abnormality of the capital physis as one probable underlying cause for a nonspherical head. PMID- 15043094 TI - Anterior femoroacetabular impingement: part I. Techniques of joint preserving surgery. AB - The exact pathomechanism responsible for osteoarthritis (OA) of the nondysplastic hip has remained unknown for many years. There is, however, emerging clinical evidence implicating femoroacetabular impingement as an etiologic factor for having early OA of the hip develop. Femoroacetabular impingement is an abutment conflict occurring between the proximal femur and the acetabular rim arising from morphologic abnormalities affecting the acetabulum or the proximal femur, or both. The repetitive mechanical conflict occurring during motion, particularly flexion and internal rotation, can lead to lesions of acetabular labrum and, even more serious, the adjacent acetabular cartilage. Surgical treatment of femoroacetabular impingement focuses on improving the clearance for hip motion and alleviation of femoral abutment against the acetabular rim. We will describe the rationale for the treatment of this condition, and discuss the technique of joint-preserving surgery. PMID- 15043095 TI - Anterior femoroacetabular impingement: part II. Midterm results of surgical treatment. AB - Femoroacetabular impingement has been shown to cause labral and chondral lesions and leads to osteoarthrosis of the hip. With the elimination of the pathogenic cause we hope to prevent or delay additional degeneration of the hip. Fourteen men and five women with a mean age of 36 years (range, 21-52 years) were treated with a surgical dislocation and offset creation of the hip. The followup averaged 4.7 years (range, 4-5.2 years). Using the Merle d'Aubigne hip score, 13 hips were rated excellent to good, with the pain score improving from 2.9 points to 5.1 points at the latest followup. There was no avascular necrosis of the femoral head. Five of the 19 patients, two with Grade 2 osteoarthrosis, two with Grade 1 osteoarthrosis but severe acetabular cartilage damage, and one with untreated ossified labrum had subsequent total hip arthroplasty (THA). In the stable hips without subluxation of the head into the acetabular cartilage defect, no additional joint space narrowing occurred. Surgical dislocation with correction of femoroacetabular impingement yields good results in patients with early degenerative changes not exceeding Grade 1 osteoarthrosis. This procedure is not suitable for patients with advanced degenerative changes and extensive articular cartilage damage. PMID- 15043096 TI - Magnetic resonance arthrography of labral disorders in hips with dysplasia and impingement. AB - Despite the fact that classic studies on osteoarthritis of the hip have shown the periphery of the hip to be prone to degeneration, it was not until recently that an abnormal acetabular labrum has been associated with osteoarthritis. This study was designed to determine whether magnetic resonance arthrography can show differences in disorders of the labrum (tears, size, ganglion formation) expected in symptomatic patients with developmental dysplasia of the hip and anterior femoroacetabular impingement. Fourteen patients in each group were evaluated preoperatively not only clinically but also with conventional radiographs and magnetic resonance arthrographs. In both conditions, disorders of the labrum localized identically with a predilection to the anterosuperior quadrant of the acetabulum. Labral tears were observed in nine hips of each group. The labrum was enlarged in 12 hips with dysplasia but in none of the hips with impingement. Ganglion formation in the periacetabular area was seen in 10 hips with dysplasia and three hips with impingement. These findings provide evidence that the anterosuperior acetabulum represents the initial fatiguing site of the hip under both conditions. Based on these data, the size of the labrum and the presence of soft tissue ganglia seem to be good predictors for the presence of developmental dysplasia, whereas the presence of tears did not differentiate between conditions. The capability of magnetic resonance arthrography to show these differences in labral disorders suggests this method is a helpful diagnostic tool that can aid in defining the most appropriate treatment strategy. PMID- 15043097 TI - Long-term survivorship of hip shelf arthroplasty and Chiari osteotomy in adults. AB - The current authors retrospectively assessed 56 hip shelf arthroplasties (48 patients) with a mean followup of 17 years (range, 15-30 years) and 89 Chiari osteotomies (82 patients) with a mean followup of 13 years (range, 6-25 years) done in adults with painful hip dysplasia. Preoperative joint space narrowing was observed in 32 of 56 shelf arthroplasties and in 67 of 89 Chiari osteotomies. Survival rates, using hip replacement as the end point, were 37% (20% to 54%) at 20 years for shelf arthroplasty and 68% (54% to 81%) at 18 years for Chiari osteotomy. The severity of preoperative arthrosis was the main factor that impaired the survivorship of shelf arthroplasty and Chiari osteotomy. With arthritic changes without joint space narrowing, the 18-year survival rates were 83% (69% to 97%) for shelf arthroplasty and 94% (89% to 99%) for Chiari osteotomy. Shelf arthroplasty is best indicated for moderate dysplasia (center edge angle >0 degrees) without severe arthrosis. Chiari osteotomy is best suited for severe dysplasia (center edge angle <0 degrees) especially without or with slight arthrosis. Chiari osteotomy also can be a salvage procedure when marked joint space narrowing is present but only if it is related to severe dysplasia (center edge angle <0 degrees). PMID- 15043098 TI - Risk factors affecting outcome of metal-on-metal surface arthroplasty of the hip. AB - We evaluated radiologic and clinical features affecting the outcome of hybrid metal-on-metal surface arthroplasty of the hip in 119 hips in patients 40 years and younger. Only the hips that had either failed or had minimum 2-year followup were reviewed. Ninety-four hips in 83 patients with a mean age of 34.2 years (range, 15-40 years) were reviewed. Seventy-one percent of the patients were males and 29% of the patients were females; 14% had previous surgery. The Chandler index and surface arthroplasty risk index were calculated. The mean followup at 3 years (range, 2-5 years) showed that three hips were converted to a total hip replacement at a mean of 27 months (range, 2-50 months) after the original surgery, and 10 hips had significant radiologic changes. The mean surface arthroplasty risk index for these 13 problematic hips versus the remaining hips was significantly higher, 4.7 and 2.6, respectively. The mean angle between the prosthesis stem and femoral shaft in the problematic group was significantly smaller than in the remaining hips: 133 degrees and 139 degrees, respectively. With a surface arthroplasty risk index score greater than 3 the relative risk of early problems is 12 times greater than if surface arthroplasty risk index less than or equal to 3. PMID- 15043099 TI - Patient factors and wear in total hip arthroplasty. AB - Polyethylene wear in vivo is multifactorial including numerous patient factors, such as gender, age, and activity. There are limitations in the clinical assessment of some of these factors and, for this reason, surrogate variables are used. For example, patient age and diagnosis frequently are used as surrogates for patient activity. The most variable and difficult factor to quantify is the frequency and intensity of use of the prosthesis during the service life of the arthroplasty. The clinical evaluation of patients with a total hip arthroplasty would be enhanced by a standardized assessment of the frequency and intensity of patient activity as it relates to the use of the prosthesis. PMID- 15043100 TI - Biologic and tribologic considerations of alternative bearing surfaces. AB - Patients who are young or active or both who require total joint replacement pose a unique challenge; their high activity demands wear-resistant bearings that will perform for decades, without suffering from the adverse effects of accumulated wear products. We discuss the tribologic and biologic properties of newly introduced bearing materials for hip prostheses. The new PEs are intended to address the aseptic loosening problem by reducing the volume of submicron PE particles to a level well below that historically associated with osteolysis. However, choosing among the several variations of the cross-linked thermally stabilized PEs is confounded by conflicting opinions regarding the optimum balance between long-term wear resistance and mechanical strength, and regarding potential effects of differences in morphologic features of the submicron-sized wear particles on their relative osteolytic potential. Metal-on-metal bearings have clinically proven wear resistance and the advantage of self-polishing, but the long-term biologic effects of metallic ions remain unknown. Ceramic-on ceramic bearings have the advantage of high biocompatibility and usually very low wear, but fracture remains a rare but catastrophic complication. The choice of an appropriate bearing couple should be made after a thorough consideration of the relative risks and potential benefits of each of these materials. PMID- 15043101 TI - Long-term results of Charnley-Kerboull hip arthroplasty in patients younger than 50 years. AB - Two hundred eighty-seven Charnley-Kerboull low friction total hip arthroplasties were done between 1975 and 1990 in 222 patients younger than 50 years. The average age of the patients at the time of the index procedure was 40.1 years. The mean followup of the entire series was 14.5 +/- 5.1 years with a median of 13.9 years. Forty-five patients (52 hips) had a followup greater than 20 years. The mean preoperative Merle d'Aubigne hip functional score was 9.6 +/- 2.5 points versus 17.2 +/- 0.8 points at the latest followup. Twenty-five revisions were documented in this series; 17 were done for aseptic loosening. The mean wear rate was 0.12 +/- 0.21 mm per year. Considering 0.1 mm per year as the threshold for a normal wear rate, 196 hips had a normal or below normal wear rate (mean, 0.02 mm per year), whereas the remaining 91 hips had an abnormally high wear rate (mean, 0.28 mm per year). The overall survival rate at 20 years was 85.4% +/- 5% using revision of either component as the end point. The only predictive factor of loosening was a wear rate higher than 0.1 mm per year. PMID- 15043102 TI - Total hip arthroplasty in patients 50 years and younger. AB - Because the young patient with a total hip arthroplasty is presumed to place increased demands on a reconstruction for a longer period than the average patient with a hip replacement, long-term results are expected to be inferior. To determine the efficacy of total hip replacements in this population, the current authors reviewed the long-term results of patients who were 50 years and younger who had cementless total hip arthroplasties at their institution, and reviewed the literature on total hip arthroplasty in younger patients. The results from the study population and the literature were encouraging. At the author's institute, during the past 20 years, 561 hip replacements were done on 488 patients in this age group, using extensively porous-coated cobalt-chromium stems matched with beaded, press-fit acetabular components of cobalt-chrome or titanium. Using the Kaplan-Meier technique, the survival rates for femoral and acetabular components, using any revision as an end point, were 89% at 10-year followup and 60% at 15-year followup. A subset of the authors' patients who were 40 years and younger (256 hips, 223 patients) had slightly inferior results, with 85% 10-year survivorship and 54% 15-year survivorship, using any revision as an end point. A comprehensive literature review also showed that long-term success can be achieved with cemented or cementless total hip arthroplasties in young patients. Because some reconstructions exhibited inferior results in younger patients, the authors recommend that surgeons be much more critical of the components used in these patients and allow long-term data to guide their decisions. PMID- 15043103 TI - Hip arthrodesis: a procedure for the new millennium? AB - Advanced hip arthrosis in the young patient represents a difficult problem. Using currently available techniques, total hip arthroplasty (THA) in this patient population likely would result in multiple revision surgeries, usually at the expense of bone in the proximal femur and acetabulum. For noninflammatory, monoarticular hip arthritis, arthrodesis remains an excellent reconstruction option. Arthrodesis can provide pain relief and return to function in the majority of patients indicated for the procedure. Current techniques provide acceptable fusion rates. If possible, techniques that protect the abductor mechanism and limit deformity of the pelvis should be used. Patient education is an important part of the treatment process. The position of the limb is important to early satisfaction and durability of the fusion. Long-term difficulties after hip fusion are related to pain in contiguous joints. Conversion arthroplasty, if necessary, can result in favorable outcomes. Familiarity with the techniques and outcomes of hip arthrodesis are important for the hip surgeon to understand. PMID- 15043104 TI - Structural properties of the axis studied in cadaveric specimens. AB - Peripheral quantitative computed tomography scans of the axis in 20 fresh cadaveric specimens obtained at autopsy were studied to investigate the internal structural properties of the axis. In all specimens, serial scans were done in three planes. The area of the dens consisted of dense cortical and trabecular bone (Region 1, above the base of the dens) and showed statistically significantly greater peripheral quantitative computed tomography values when compared with the anterior part of the body of the axis (Region 2, below the base of the dens). An age-related progressive decrease of peripheral quantitative computed tomography values was observed, with specimens from subjects younger than 40 years showing statistically significantly greater peripheral quantitative computed tomography values when compared with those from subjects older than 40 years. A small area of trabecular bone with decreased density was found close to the anteroinferior edge of the body of the axis in the specimens from younger subjects. In contrast, a large area of trabecular bone with decreased density extending anteroinferiorly to superoposteriorly to the base of the dens was found in the specimens from older subjects. Based on these findings, we think that dens fracture classification systems and fixation techniques should be reconsidered. PMID- 15043105 TI - Anatomy of the posterior iliac crest as a reference to sacral bar insertion. AB - There are no detailed anatomic studies focusing on the posterior iliac crest although it frequently is used for posterior stabilization of unstable pelvic fractures. Anatomic dissections were done to evaluate the size of the extraarticular region of the posterior iliac crest and its relationship to the lumbosacral lamina and to show on cadavers the level of sacral bar placement that offers safe and solid fixation. Sixty cadavers were dissected bilaterally. Fifty one were male and nine were female. The distance between the posterior wall of the sacral canal and the tip of the iliac crest was measured at various levels between the level of the upper border of L5 lamina to the level of the posterosuperior iliac spine. In all the dissections the greatest distances were at the level of the L5-S1 junction, which consequently is the safest level for good bony purchase. The entire length of the posterior iliac crest from the level of the upper border of L5 lamina to the posterosuperior iliac spine was shown to be appropriate for safe and solid bar fixation because all of the distance measurements were greater than 13 mm, which is the smallest safe distance. Below the posterosuperior iliac spine level, insertion of the sacral bars was dangerous because the average measured distance was only 10.38 mm. PMID- 15043106 TI - Sexual activity, pregnancy, and childbirth after periacetabular osteotomy. AB - The purpose of the current study was to determine the effect periacetabular osteotomy has on sexual activity, pregnancy, and childbirth. Eighty-eight female patients who had periacetabular osteotomy were asked to complete a questionnaire concerning the presence of hip pain and changes in the frequency, positions, and satisfaction of sexual activity after periacetabular osteotomy. They also were asked about hip pain during pregnancy and after childbirth, type of delivery, and complications during childbirth. Eighty-four patients (89%) either were very satisfied or satisfied with the surgery. Thirty-eight (40%), 44 (46%), and 24 (25%) patients reported changes in the frequency, positions, and satisfaction of sexual intercourse, respectively. Sixteen women had 24 pregnancies. Fifteen women had vaginal deliveries and eight had Cesarean sections. Less than 5-mm medialization of hip center after correction was associated with pain (in seven of seven patients) and decreased range of motion (in eight of nine patients) during sexual activity. Acetabular retroversion was found in 16 patients postoperatively, and 15 of them had hip pain after periacetabular osteotomy. None of the patients who reported an increased frequency of sexual intercourse had retroversion. The majority will be able to deliver a child vaginally. PMID- 15043107 TI - Early osteolysis associated with trunion-liner impingement. AB - Osteolysis is perhaps the biggest unsolved problem in joint arthroplasty. Many factors contribute to osteolysis, some of which cannot be controlled. It is imperative to minimize controllable factors contributing to osteolysis whenever possible. We report four cases of trunion impingement on an elevated rim liner, which may have contributed to early aggressive osteolysis. Although the use of an elevated liner may result in a lower incidence of postoperative instability, the trade-off in using such a device must be understood. PMID- 15043108 TI - A quantitative anatomic characterization of the quadrilateral surface for periacetabular osteotomy. AB - The periacetabular osteotomy described by Ganz et al is used widely, and includes an outward osteotomy from the quadrilateral surface. Because intraarticular extension of the osteotomy can complicate the Ganz osteotomy, it is important to image the margin of the hip. To prevent this complication, and to do this procedure more safely, 32 hemipelves from cadavers were used in the current study. Some landmarks were selected that can be clarified on the quadrilateral surface during the periacetabular osteotomy. The acetabulum was hollowed out using an acetabular reamer of the same size as each femoral head, and the margin of the penetrated hole through the acetabulum was determined using these landmarks. The posterior margin of the hip is located approximately 2 cm anterior to the sciatic notch. The anatomic guidepoint for the osteotomy of the ischium averaged 14 mm inferior to the distal margin of the hip. By clarifying the margin of the hip presumed on the quadrilateral surface in this way, the periacetabular osteotomy can be done more safely, without causing complications such as intraarticular chisel penetration. PMID- 15043109 TI - A posterior approach to primary total hip arthroplasty with soft tissue repair. AB - To determine whether repair of the posterior soft tissue structures affects dislocation rate, a comparison of the posterolateral approach with and without a posterior soft tissue repair in primary total hip arthroplasty was done. Between January 1993 and December 1998, 250 consecutive primary total hip arthroplasties (220 patients) without a posterior soft tissue repair followed by 96 consecutive primary total hip arthroplasties (83 patients) with a posterior soft tissue repair in the posterolateral approach were done. With the exception of a posterior soft tissue repair, all surgical procedures were done in the same manner. A dislocation rate of 6.4% in 250 hips without a posterior soft tissue repair in the posterolateral approach was reduced to 1% in 96 hips with a posterior soft tissue repair. The results were statistically significant. The current results indicate that the posterior structures should be preserved as much as possible and repaired to reduce dislocation in the posterolateral approach. PMID- 15043110 TI - Nerve injury and limb lengthening after hip replacement: treatment by shortening. AB - Nineteen patients had a severe neurologic deficit and persistent dysesthetic pain after total hip replacement. During surgery their limbs had been lengthened by 1.3 to 4.1 cm. Patients with other reasons for nerve injury such as hematoma, dislocation, or direct operative trauma were excluded. Also excluded were patients whose lengthening was part of the operative plan to address previous shortening. None of the patients had neurologic deficits or dysesthetic pain preoperatively. Patients were offered revision hip surgery to shorten the limb. Seventeen patients proceeded to surgery. Two acetabular revisions, five modular femoral head exchanges, and 10 femoral component revisions were done. The average shortening was 1.5 cm (range, 0.5-3.6 cm). The trochanter was advanced in four patients and a constrained acetabular liner was used in four patients. Nine hips were stable without additional measures. There were no complications and nine patients had an excellent result. This was defined as elimination of their dysesthetic pain. Two patients had partial improvement and six patients had no improvement. Seven of 11 patients with motor deficits had improvement in strength with three making a full recovery. When painful neurologic symptoms accompany limb lengthening after total hip replacement, revision surgery may be helpful, although patients should be informed the rate of success is far from uniform. PMID- 15043111 TI - Late loosening of press-fit cementless acetabular components. AB - Between January 1987 and September 1990, 67 consecutive cementless total hip arthroplasties were implanted in 59 patients by one surgeon. The mean age of the patients at surgery was 57 years (range, 23-80 years). All acetabular components were plasma-sprayed titanium hemispheric cups with four peripheral rim fins, but additional screw fixation was not used. The mean followup was 10.4 years (range, 8.8-12.5 years). With revision as the endpoint, the failure rate of this acetabular component at a mean of 10.4 years was 28% (19/67). Of the 56 patients (56 hips) with radiographic followup, loosening of the acetabular shell occurred in 10 hips in 10 patients (18%). Seventy percent of these loosened cups failed by tilt which occurred in a rapid manner; all of the patients required revision surgery. We examined the manner of loosening of a press-fit acetabular component after early (5-year) results showed high hip scores and a low failure rate. PMID- 15043112 TI - Total knee arthroplasty for patients 90 years of age and older. AB - This study was done to review the reliability, durability, and safety of primary and revision total knee arthroplasty for patients 90 years of age and older. Because little data exist about the safety or efficacy of total knee replacement in this elderly population some physicians may be hesitant to recommend elective surgery to patients of this age. Fifty-one total knee replacements were done for 41 patients age 90 to 102 years (mean, 92 years). Forty-four primary total knee replacements and seven revision total knee replacements were done. Each patient was followed up until death or for a minimum of 2 years (mean, 4.0 years). The Knee Society pain scores improved significantly for the primary and revision groups from preoperative mean scores of 30 and 29 points to latest followup scores of 86 and 87 points, respectively. One patient died in the early postoperative period of causes related to the operation. The remaining patients have lived an average of 4.4 +/- 2.2 years after the primary knee arthroplasties and 5.7 +/- 2.8 years after the revision knee arthroplasties. No knees have required revision subsequently. Total knee replacement was reliable, durable, and safe in this group of patients older than 90 years. Primary care physicians and surgeons should be aware that primary and revision total knee replacement can be done safely and effectively for patients 90 years and older and result in years of marked pain relief for those patients. PMID- 15043113 TI - In vivo kinematics for fixed and mobile-bearing posterior stabilized knee prostheses. AB - This is the first in vivo kinematic study to compare mobile-bearing with fixed bearing prostheses in patients who had total knee arthroplasties. Femorotibial contact positions for 40 patients implanted with either a fixed-bearing or mobile bearing prosthesis were analyzed using videofluoroscopy. Femorotibial contact paths were determined using a computer automated model-fitting technique. Nineteen of 20 patients in each group experienced posterior femoral rollback of their lateral condyles, with a mean of 3.6 and 3.7 mm for fixed-bearing and mobile-bearing prostheses respectively. Eighteen patients who had mobile-bearing prostheses and 17 patients with fixed-bearing knee prostheses experienced a normal pattern of axial rotation of 7.3 degrees and 4.1 degrees respectively. Eleven of 20 (55%) patients who had mobile-bearing prostheses implanted and eight of 20 (40%) patients who had fixed-bearing prostheses implanted did not experience femoral condylar lift-off. The remaining knees had condylar lift-off less than 2.4 mm for fixed-bearing prostheses and 1.7 mm for mobile-bearing prostheses, respectively. Patients who had mobile-bearing prostheses implanted experienced greater axial rotation and less condylar lift-off than patients who had fixed-bearing prostheses implanted. Both cruciate ligaments are sacrificed for the mobile and fixed-bearing total knee replacements. The results from the current study showed that, in both groups, the majority of patients experienced kinematics similar to those of a normal knee. However, the extent of lateral femoral condyle posterior rollback and the extent of axial rotation were less. PMID- 15043114 TI - Hemangioendothelioma of the spine. AB - A review of the patient files from our institution's oncology department showed that between 1950 and 2000, 11 patients were diagnosed with hemangioendothelioma of the spine, a rare, low-grade malignant vascular tumor. After reevaluation of the diagnoses by orthopaedic oncologists and pathologists, three patients were excluded; therefore eight patients formed the study group. The eight lesions were located in the thoracic (three) or lumbar spine (five, one in each vertebral level). Computed tomography scans revealed expansile lytic process. All lesions involved the vertebral body, but only one was diffuse with spinous process involvement. Treatment included: external beam irradiation alone (one patient), curettage and external beam irradiation (one patient), laminectomy and external beam irradiation (two patients), anterior resection only (two patients), and anterior resection with postoperative external beam irradiation (two patients). Patients without spinal stabilization had chronic low back pain; no patient with spinal stabilization had such pain. One patient who had surgical resection and radiation therapy had a radiation-induced sarcoma develop after 4 years. One patient who was treated with radiation therapy alone had a metastatic lung lesion develop. No other patient had tumor recurrence or progression. Therefore, patients with hemangioendothelioma of the spine may be treated with radiation therapy alone, surgery alone, or a combination thereof. However, because the number of patients in the current series was small, no definitive recommendations regarding treatment may be made. Resection of large lesions and stabilization of vertebral collapse may decrease back pain. PMID- 15043115 TI - Synovial sarcoma after chemotherapy for osteosarcoma: a case report. AB - A 23-year-old man had eight cycles of adjuvant chemotherapy including doxorubicin, cisplatin, methotrexate, and ifosfamide and radical surgery for biopsy-proved osteosarcoma of the right fibular shaft. Two years after the initial diagnosis, he noticed a mass in the medial aspect of his right knee. Magnetic resonance imaging scans revealed a soft tissue tumor measuring 2 x 2 cm in the pericapsular region of the right knee. Histologically, this soft tissue tumor was composed of spindle cells with occasional atypical mitoses and without matrix formation. Immunohistochemically, the tumor cells were positive for vimentin, cytokeratin, and epithelial membrane antigen, and negative for alpha smooth muscle actin. A fusion gene, SYT-SSX was detected with reverse transcription-polymerase chain reaction. From the results, the secondary tumor was diagnosed as a synovial sarcoma. The current case of double sarcomas is rare. Both sarcomas were diagnosed accurately using immunohistochemical and molecular procedures. This case suggests a positive association between a second tumor and chemotherapy including intraarterial perfusion of doxorubicin. PMID- 15043116 TI - Paget's disease of bone in patients younger than 40 years. AB - Paget's disease of bone, although common in the United States, is relatively rare in patients younger than 40 years. In a large archival series, 10% of patients with Paget's disease of bone were younger than 40 years. Pain followed by pathologic fracture was the most common presenting complaint in this group of patients. There was no gender difference comparing the younger group with the older group of patients. Of the 521 patients in this series, only 10 were African American; the others were Caucasians. Seven African-American patients were in the group of patients who were younger than 40 years. Appendicular and monostotic bone involvement was more frequent in the younger age group although these differences were not statistically significant. The incidence of malignant transformation in pagetic bone was high (30.3%), and is a reflection of the high volume of uncommon tumor referrals to our consultation service. However, there were no cases of associated malignancies in the group of patients younger than 40 years. PMID- 15043117 TI - Aspirin therapy and bleeding during proximal femoral fracture surgery. AB - To assess the effect of daily low-dose aspirin therapy on perioperative bleeding of patients operated on for proximal femoral fracture, we did a prospective case control study. During 14 months, we followed up 104 patients, 39 of whom were taking aspirin before the injury. The bleeding was estimated by the number of blood units needed perioperatively, the change in hemoglobin values, and followup on complications and drain volume. The aspirin-treated group received an average of 0.5 units of blood more than the control group, postoperatively. This finding was statistically significant. The groups did not differ significantly in any other bleeding parameter. No major bleeding occurred in the patients. It is safe to do surgery for a proximal femoral fracture in patients who are taking aspirin. PMID- 15043118 TI - Pseudoaneurysm after tibial nailing. AB - This is a case report of a patient with a pseudoaneurysm of the anterior tibial artery after lateral to medial distal locking of an intramedullary nail for a tibia shaft fracture. PMID- 15043119 TI - The role of soft tissues in plate fixation of proximal phalanx fractures. AB - The tension band effect of plate fixation and the contribution of soft tissues to that effect was examined biomechanically in human proximal phalanges. Forty-six proximal phalanges in whole cadaver hands with all soft tissues in place (intact) and 43 proximal phalanges stripped of soft tissues (denuded) were tested. After midshaft osteotomy, each proximal phalanx was fixed internally with a dorsal minicondylar plate, a lateral minicondylar plate, a dorsal straight plate, or a lateral straight plate. Specimens were tested in three-point apex dorsal bending to clinical failure, defined as 30 degrees angulation. Ultimate moment (stability) at this angulation was similar among the four fixation methods in the specimens with all soft tissues intact. Stability also was similar among these methods in the denuded specimens. There were no significant differences in stability between minicondylar and straight plates or between dorsal and lateral plates in the specimens with soft tissues, nor were there significant differences between these groups in the denuded specimens. The stability of the four fixation methods was significantly greater in the specimens with soft tissues than in the denuded specimens. Soft tissues increased the stability of lateral minicondylar plates by 163%, lateral straight plates by 157%, dorsal minicondylar plates by 126%, and dorsal straight plates by 104%, providing a dorsal tension band effect that counteracted the buttress (compression) of the volar fracture surfaces of the phalanx. The results suggest that in the clinical setting a laterally placed straight or minicondylar plate may provide as much stability to a phalanx with a midshaft fracture as does the traditional, more invasive dorsally placed minicondylar or straight plate. These findings must be evaluated with caution, however, because all specimens were from embalmed cadavers, and the formalin fixation may have augmented the stability and stiffness of the soft tissues in the intact specimens. A subsequent pilot study comparing intact proximal phalangeal specimens that were formalin-fixed with those that were fresh-frozen showed a significant increase in stability and stiffness of formalin-fixed specimens. PMID- 15043120 TI - Septic arthritis in a patient with juvenile rheumatoid arthritis. AB - The acute flare of joint inflammation in the child with known juvenile rheumatoid arthritis causes concern primarily regarding the need for additional or modified medical treatment. Acute joint inflammation in an otherwise healthy child creates concern regarding the existence of joint infection. In the early phase of disease, the clinical findings and symptoms of an inflamed joint attributable to juvenile rheumatoid arthritis or infection may be similar and difficult to differentiate from the other. Juvenile rheumatoid arthritis usually is well controlled by medical interventions, however, the initiation of specific treatment is more urgent in children with joint sepsis. The following case report is presented to emphasize the difficulty in evaluation of patients with known juvenile rheumatoid arthritis and coexistent septic arthritis, and to discuss the methods used to differentiate between the two conditions. PMID- 15043121 TI - Infection of a total hip arthroplasty with Prevotella loeschii. AB - Infection is a serious complication of total hip replacement. It has been proposed that 6% of all infections after total hip arthroplasty may be of dental origin through hematogenous spread. However, no conclusive evidence that the mouth is a definitive source for infection of a total hip replacement has been reported. In the current case, Prevotella loeschii, a pigmented bacteroides species was identified in a total hip replacement. Prevotella loeschii is an organism which exclusively inhabits the dental region. Hematogenous spread of Prevotella loeschii may occur after penetration of the mucosal barrier in cases of endodontic or periodontic lesions, pericoronitis, or complications of tooth extraction. The involvement of Prevotella loescheii in an infection in a patient who had a total hip arthroplasty is strong evidence for the mechanism of a hematogenous infection from a dental source. PMID- 15043122 TI - Bioabsorbable expansion bolt fixation in anterior cruciate ligament reconstruction. AB - The current study evaluated initial fixation strength of a bioabsorbable expansion bolt compared with interference screw fixation in anterior cruciate ligament reconstruction using a bone-patellar tendon-bone graft. Thirty calf tibial plateaus with adjacent patella and extensor ligaments were used. Bioabsorbable poly-L-lactide interference screws were used for graft fixation in Group I, titanium screws in Group II, and bioabsorbable poly-DL-lactide expansion bolts were used in Group III. The mean force-to-failure (+/- standard deviation) in the three groups was 487 +/- 205 N, 713 +/- 218 N, and 594 +/- 224 N, respectively. The differences between Groups I and II were significant. No statistical differences were found regarding stiffness. Graft damage was significantly less in Group III compared with screw fixation. The fixation concept of an expansion bolt shows similar fixation strength and less graft damage compared with the established interference screw fixation. Because of the total absence of torque forces in contrast to bioabsorbable screws, the risk of implant breakage is minimized. PMID- 15043123 TI - Distraction osteogenesis (callotasis) for pelvic closure in bladder exstrophy. AB - The success of urogenital reconstruction in bladder exstrophy is dependent on how successfully the pelvic ring can be closed. In patients with this disorder, the pubic bones are short and separated. A two-component plate-rod combination was designed to widen the pelvic ring using distraction osteogenesis (callotasis). After a middiagonal iliac osteotomy, two components of the implant are connected with two bolts passing through the iliac bone. After 10 days, the rods outside the skin are approximated gradually. When the pubic bones get close, they are fixed by heavy nonabsorbable sutures. The current study included 14 patients who were operated on between 1990 and 1996. The mean followup was 6 years. No neurologic or vascular complications developed. Successful tension-free closure of the abdominal wall was achieved in all but one patient. Urogenital reconstruction was done in the second stage. In all but one patient, the bladder was closed in one stage of urogenital reconstruction. In one patient (7%), a deep infection developed for which the patient needed early implant removal resulting in failure of the urogenital reconstruction. The described method addresses the pathoanatomy of the pelvic ring. The diameter of the pelvic ring can be widened and the pubic bones can be approximated without a significant increase in tension. PMID- 15043124 TI - Intraoperative measurement of force-length relationship of human forearm muscle. AB - The specific relationship between force and length is one of the most important characteristics of vertebrate muscle. The only accurate method to measure the force-length characteristics is to generate a set of isometric force-time plots at different muscle lengths. In humans, such force-length characteristics mostly are based on indirect measurements that have their limitations. A method of direct, in vivo measurement of force-length characteristics of the human flexor carpi ulnaris muscle using relatively simple equipment during transposition surgery is presented. The method is proven reproducible, with an overall estimated error of 2.8%. PMID- 15043125 TI - Depot local anesthetic in polymethylmethacrylate bone cement: a preliminary study. AB - Acrylic bone cement has been used successfully as a slow-release depot for antibiotics after orthopaedic surgery. The feasibility of administering local anesthetics in this way was examined in this preliminary in vitro study. Discs weighing approximately 4 g were prepared from five brands of acrylic cement (40 g) containing as much as 2 g of anesthetic base. Elution of the anesthetics into saline was measured during 72 hours. Prilocaine eluted the fastest and bupivacaine the slowest, with lidocaine between them. The elution rates were greatest in the first hour, declining thereafter. Rates also depended on the brand of cement with the quickest elution from CMW3 and the slowest from Surgical Simplex P. Using cement as a depot, therapeutic levels of a drug should be achievable in vivo at a negligible risk of toxicity. Before in vivo trials it is necessary to optimize elution of drugs in relation to the cement (brand, microstructure, method of preparation) and the concentration of a drug in the cement above which the cements' mechanical and adhesive properties are compromised. PMID- 15043126 TI - The effects of storage on fresh human osteochondral allografts. AB - Historically, fresh human osteochondral allografts have been stored in lactated Ringer's solution at 4 degrees C and then transplanted as quickly as possible, generally within 2 to 5 days, to ensure delivery of a high level of viable chondrocytes. Recently, allograft distribution companies have begun to provide fresh osteochondral allografts that are stored in a proprietary culture medium usually for at least 2 weeks before delivery to the surgeon for implantation. The effects of such storage on human cartilage have not been well-defined. In the current study the effects of storage in lactated Ringer's solution and in culture media were assessed. After 7 days of storage in lactated Ringer's solution, a significant decline in chondrocyte viability and metabolic activity was seen. Culture media provided significantly better preservation of the cartilage with viability and metabolic activity remaining essentially unchanged from baseline for as many as 14 days. The biochemical and biomechanical properties of the extracellular matrix remained stable with storage in both solutions with time. These data suggest that osteochondral allografts stored under traditional conditions in lactated Ringer's solution should continue to be implanted as quickly as possible and certainly within 7 days of donor death. If kept in culture media, the storage duration may be extended to approximately 2 weeks. PMID- 15043127 TI - Low intensity pulsed ultrasound stimulates osteogenic activity of human periosteal cells. AB - The effect of low intensity pulsed ultrasound on human periosteal cells was investigated. Normal human periosteum was obtained to culture the periosteal cells. After characterization, cultures of periosteal cells at Days 2 and 4 were treated with ultrasound for 5, 10, and 20 minutes respectively. Assessments were done to assess total number of viable cells, cell proliferation, alkaline phosphatase activity, osteocalcin secretion, vascular endothelial growth factor expression, and calcium nodule formation. With the cells not treated with ultrasound as the control, the results showed that ultrasound did not affect the total number of viable cells. It stimulated cell proliferation at the early phase of cell culture. The activity of alkaline phosphatase was increased significantly in the culture at Day 4. A similar effect was seen with osteocalcin secretion and the responses were dose-dependent. The vascular endothelial growth factor secretion increased in Day 2 and Day 4 cultures with the dose-dependent effect. Formation of calcium nodules was significantly higher with ultrasound treatment. We think that low intensity pulsed ultrasound stimulated periosteal cell proliferation and differentiation toward osteogenic lineage. The dose-dependent effect on osteogenic activities may modify the existing treatment regimen. Ultrasound treatment should be started from the beginning of fracture healing. PMID- 15043128 TI - Tourniquet-induced ischemia and reperfusion in human skeletal muscle. AB - Microdialysis conceivably enables longitudinal and simultaneous investigation of several metabolites by repeated measurements in skeletal muscle. We used and evaluated microdialysis as an in vivo method to characterize the time-course and relative kinetics of pyruvate, glucose, lactate, glycerol, hypoxanthine, uric acid, and urea, in skeletal muscles, exposed to ischemia and reperfusion, in eight patients having arthroscopic-assisted anterior cruciate ligament reconstruction. A dialysis probe was implanted before surgery in the rectus femoris muscle. Dialysate samples were collected at 10-minute intervals at a flow rate of 1 microL/minute until 2 hours after tourniquet deflation. Ninety minutes of ischemia resulted in accumulation of lactate (234% +/- 38%), hypoxanthine (582% +/- 166%), and glycerol (146% +/- 46%), consumption of glucose (54% +/- 9%) and pyruvate (16% +/- 44%), and a slight decrease of urea (78% +/- 11%) compared with baseline (100%). Uric acid was unchanged (95% +/- 12%). Within 90 minutes after tourniquet deflation the concentrations were virtually normalized for all measured metabolites, suggesting that the duration of ischemia was well tolerated by the patients. The results indicate that the use of microdialysis for monitoring energy metabolic events during orthopaedic surgery that requires ischemia and reperfusion is feasible and safe. PMID- 15043129 TI - Lesion of the right proximal tibia in a 12-year-old boy. PMID- 15043130 TI - Elbow mass in a 2-year-old girl. PMID- 15043131 TI - Lower leg pain in a 17-year-old boy. PMID- 15043132 TI - Synthesis and antiviral evaluation of some 3'-fluoro bicyclic nucleoside analogues. AB - The synthesis of 3'-fluoro analogues of recently discovered highly potent anti VZV furanopyrimidine deoxynucleosides (BCNAs) is herein reported, for both the alkyl and alkylphenyl series. The compounds are tested against a range of herpes viruses and display poor activity, strongly supporting the notion of the importance of the presence of a 3'-OH for antiviral activity. PMID- 15043133 TI - Synthesis and antiviral activity of novel fluorinated 2',3'-dideoxynucleosides. AB - A series of 5-(trifluoroethoxymethyl)-2',3'-dideoxyuridines and 5 [bis(trifluoroethoxy)-methyl]-2',3'-dideoxyuridines have been prepared and screened for antiviral activity. The conformations of these compounds are discussed on the bases of NOE studies and the MO calculations. Modelling and NOE studies suggest both syn- and anti conformations for these 5-(2,2,2 trifluoroethoxymethyl)- and 5-[bis(2,2,2-trifluoroethoxy)-methyl]- derivatives. The NOE parameters are also suggested to be more attributable to the nature of the fluorine atom than to structural or conformational changes. Compounds 17, 26 and 30 showed some activity in anti-HIV-1 and anti-HIV-2 assays, but the compounds were devoid of activity against HSV and human rhinovirus. The compounds tested exhibited low cytotoxicity and were inactive against a bank of cancer cells in vitro. PMID- 15043134 TI - An improved total synthesis of triciribine: a tricyclic nucleoside with antineoplastic and antiviral properties. AB - We describe an efficient total synthesis of triciribine, a tricyclic nucleoside with antineoplastic and antiviral properties, starting from 4-amino-6-bromo-5 cyanopyrrolo[2,3-d]pyrimidine. PMID- 15043135 TI - Tandem mass spectrometry for structure assignments of wye nucleosides from transfer RNA. AB - The tricyclic wye nucleoside family of eight known members constitutes one of the most complex and interesting series of posttranscriptionally modified nucleosides in transfer RNA. The principal reaction paths represented in collision-induced dissociation mass spectra of wye bases and their analogs have been studied in order to determine those structural features that can be readily established by mass spectrometry. The main routes of fragmentation are determined by the presence vs. absence of an amino acid side chain at C-7 (1H-imidazo[1,2-a]purine nomenclature). The common methionine-related side chain is cleaved at two points, providing a ready means of establishing the presence and net level of side chain modification. For those molecules without a side chain, the initial reaction steps are characteristically controlled by the presence vs. absence of methyl at N-4, allowing determination of the methylation status of that site. In the latter case initial opening of the central (pyrimidine) ring, in analogy to the dissociation behavior of guanine, causes loss of identity of C-6/C-7 so that placement of a single methyl at either site is not possible. Subsequent complex reaction paths follow, which include loss of CO and sequential loss of two molecules of HCN. PMID- 15043136 TI - Nucleosides. LXV. Synthesis of new pteridine-N8-nucleosides. AB - A general synthetic approach to various isoxanthopterin-nucleosides starting from 6-methyl-2-methylthio-4(3H),7(8H)-pterdinedione (1) has been developed. Ribosylation with 1-O-acetyl-2,3,5-tri-O-benzoyl-beta-D-ribofuranose via the silyl-method led to 2 and reaction with 1-chloro-2-deoxy-3,5-di-O-p-toluoyl-alpha D-ribofuranose using the DBU-method afforded 28. Protection of the amide function at O4 by benzylation to 5 and by a Mitsunobu reaction with 2-(4 nitrophenyl)ethanol to 29 gave soluble intermediates which can be oxidized to the corresponding 2-methylsulfonyl derivatives 8 and 30, respectively. Nucleophilic displacement reactions of the highly reactive 2-methylsulfonyl functions by various amines proceeded under mild conditions to isoxanthopterin-N8-ribo- (11 17) and 2'-deoxyribomucleosides (31-33). Debenzylation can be achieve by Pd catalyzed hydrogenation (9 to 19) and cleavage of the npe-protecting group (31, 32 to 34, 35) works well with DBU by beta-elimination. PMID- 15043137 TI - L-deaza-5'-noraisteromycin. AB - (+/-)-1-Deazaaristeromycin (4) has been reported to be an inactivator of S adenosylhomocysteine (AdoHcy) hydrolase and, as a consequence, to affect S adenosylmethionine (AdoMet) mediated macromolecular biomethylations. To extend this to our program focused on 5'-noraristeromycin derivatives as inhibitors of the same hydrolase enzyme as potential antiviral agents, both enantiomers of 1 deaza-5'-noraristeromycin (5 and 20) have been prepared. Compounds 5 and 20 were evaluated against the following viruses: vaccinia, cowpox, monkeypox, Ebola, herpes simplex type 1 and 2, human cytomegalovirus, Epstein Barr, varicella zoster, hepatitis B, hepatitis C, HIV-1 and HIV-2, adenovirus type 1, measles, Pichinde, parainfluenza type 3, influenza A (H1N1 and H3N2), influenza B, Venezuelan equine encephalitis, rhinovirus type 2, respiratory syncytial, yellow fever, and West Nile. No activity was found nor was there any cytotoxicity to the viral host cells. PMID- 15043138 TI - How a modification (8-aza-3-deaza-2'-deoxyguanosine) influences the quadruplex structure of Hotoda's 6-mer TGGGAG with 5'- and 3'-end modifications. AB - We have synthesized a modified 6-mer using Hotoda's 6-mer TGGGAG with 5'- and 3' end modifications as a template. We have replaced from one to all four 2' deoxyguanosines by 8-aza-3-deaza-2'-deoxyguanosine (c3z8dG, 1) in order to investigate the anti-HIV structure activity relationship (SAR). ODN 4 (TGGG*AG) is the only one that exhibits a moderate anti-HIV-1 activity. PMID- 15043139 TI - Second generation of cycloSal-pronucleotides with esterase-cleavable sites: the "lock-in"-concept. AB - A conceptual extension of the cycloSal-pronucleotide approach is presented. The characteristic feature of the new cycloSal-derivatives of the anti-HIV active nucleoside analogue d4T 1 is the incorporation of an enzymatically cleavable carboxylic ester moiety with the intention to trap the triesters inside cells ("lock-in"-concept). CycloSal-triesters bearing different ester groups in the 3 or 5-position of the cycloSal-moiety are described. Surprisingly, only acetyl-and levulinyl esters are cleaved readily in CEM cell extracts while alkyl esters were found to be stable. Nevertheless, in in-vitro anti-HIV assays most of the compounds achieve the thymidine-kinase bypass, thus proving that they act at least as nucleotide delivery systems. PMID- 15043140 TI - Rearrangement reactions of 1,N2-isopropenoguanine cyclonucleosides. AB - Under acid-catalyzed transglycosylation conditions 5',8-cyclo-8-oxo-1,N2 isopropenoguanine nucleosides (1, 10) undergo a ring-opening reaction to 8-oxo 1,N2-isopropenoguanosine derivatives (4, 11) followed by recyclization to fluorescent 5',3-cyclonucleosides (2, 12). PMID- 15043141 TI - An efficient route to novel 4,5-di- and 2,4,5-tri substituted imidazoles from imidazo[1,5-a]-1,3,5-triazine (5,8-diaza-7,9-dideazapurine) derivatives. AB - Two new types of imidazole derivatives: N-(2-R1-5-R2-1H-imidazol-4-yl) thioureas 7a-g and N-(2-R1-5-R2-1H-imidazol-4-yl) formamides 8b,c,g were obtained in high yields by the hydrolytic degradation of 6-R1-8-R2-2-thioxo-2,3-dihydroimidazo[1,5 a]-1,3,5-triazin-4(1H)-ones 5a-g and 6-R1-8-R2-imidazo[1,5-a]-1,3,5-triazin-4(3H) ones 6b,c,d, respectively. The tautomeric preferences of the new imidazoles were determined. PMID- 15043142 TI - Functionalization of guanosine and 2'-deoxyguanosine at C6: a modified Appel process and S(N)Ar displacement of imidazole. AB - Treatment of sugar-protected 2-N-trityl derivatives of guanosine and 2' deoxyguanosine with imidazole/triphenylphosphine/iodine/ethyldiisopropylamine gives the corresponding 6-(imidazol-1-yl)-2-(tritylamino)purine nucleosides. S(N)Ar displacement of the imidazole moiety with nucleophiles provides 2-amino-6 substituted-purine nucleosides and 2'-deoxynucleosides. PMID- 15043143 TI - Biologically active antisense phosphorothioate oligodeoxyribonucleotides: synthesis, characterization, and studies of 3'-terminal phosphorothioate monoester analogues. AB - Multiple phosphorothioate oligonucleotides containing a 3'-terminal negative charge were synthesized and characterized. Influence of the added negative charge on activation of duplexes by RNase H was investigated. No additional help in recruitment of RNase H was observed. PMID- 15043144 TI - Synthesis and biological activity of 5-fluorotubercidin. AB - The electrophilic fluorination of 4-chloropyrrolo[2,3-d]pyrimidine (1) was studied culminating a 59% conversion of compound 1 to 4-chloro-5 fluoropyrrolo[2,3-d]pyrimidine (2) using Selectfluor. This transformation proceeded via the 4-chloro-5,6-dihydro-5-fluoro-6-hydroxypyrrolo[2,3-d]pyrimidine (3) in a 9:1 trans:cis ratio. The trans isomer of compound 3 was studied by 1H NMR and 19F NMR, and the 5-H tautomer (4) was observed as another intermediate. A modified Vorbruggen procedure of compound 2 and tetra-O-acetylribose gave 4 chloro-5-fluoro-7-(2,3,5,-tri-O-benzoyl-beta-D-ribofuranosyl)pyrrolo[2,3 d]pyrimidine (6) in a 65% yield. Treatment of compound 6 with ammonia (l) in dioxane gave 5-fluorotubercidin (7). No antibacterial activity was observed. An MTT assay (Promega) against Huh-7 liver cells, normal mouse spleen cells stimulated with Con A (a T-cell mitogen), and normal mouse spleen stimulated with LPS (a B-cell mitogen) showed no significant toxicity. Increased activity of 7 over tubercidin was observed against L-1210 cells and toxicity in fibroblast cells was reduced. PMID- 15043145 TI - Improved and reliable synthesis of 3'-azido-2',3'-dideoxyguanosine derivatives. AB - An improved synthesis of N2-protected-3'-azido-2',3'-dideoxyguanosine 20 and 23 is described. Deoxygenation of 2'-O-alkyl (and/or aryl) sulfonyl-5' dimethoxytritylguanosine coupled with [1,2]-hydride shift rearrangement gave protected 9-(2-deoxythreo-pentofuranosyl)guanines (10, 12 and 16). This rearrangement was accomplished in high yield with a high degree of stereoselectivity using lithium triisobutylborohydride (L-Selectride). Compounds 10, 12 and 16 were transformed into 3'-O-mesylates (18 and 21), which can be used for 3'-substitution. The 3'-azido nucleosides were obtained by treatment of 18 and 21 with lithium azide. This procedure is reproducible with a good overall yield. PMID- 15043146 TI - A new approach for the synthesis of novel 5-substituted isodeoxyuridine analogs. AB - Cyclic sulfates of carbohydrates provide excellent synthons for the preparation of isodeoxyuridines through direct nucleophilic substitution reactions. These substitution reactions have exceptional regioselectivity. The products of the reactions served as key precursors for the synthesis of 5-substituted isodeoxyuridines via the Stille and Heck coupling reactions. Interestingly, unprotected nucleosides could be used in these metal-mediated functionalizations. The methodologies are general and allow ready access to a variety of C-5 functionalized isomeric deoxyuridines, but also have the potential to be extended to other nucleoside analogs. PMID- 15043147 TI - Synthesis of uridine 5'-[2-S-pyridyl-3-thio-alpha-D-galactopyranosyl diphosphate]: precursor of UDP-thiogal sugar nucleotide donor substrate for beta 1,4-galactosyltransferase. AB - The syntheses of a novel uridine diphosphate galactose (UDP-Gal) analog, (UDP 2,4,6-tri-O-acetyl-3-S-acetyl-3-thio-alpha-D-galactopyranose) (11) and the thiolpyridine protected (Uridine 5'-[3-S-(2-S-pyridyl)-3-thio-alpha-D galactopyranosyl diphosphate) analog (12) are described. The reported synthesis relies on the novel use of thiolpyridine to generate 12 which is a suitably protected intermediate for generating a UDP-thioGal derivative by reduction prior to enzyme transfer via beta-1,4-galactosyltransferase. PMID- 15043148 TI - Intercalating nucleic acids: the influence of linker length and intercalator type on their duplex stabilities. AB - Six new examples of intercalating nucleic acids were synthesized in order to evaluate the dependence of the length of the linker between oligo and intercalator on the thermal stability of their corresponding duplexes and triplexes. PMID- 15043149 TI - Regioselective synthesis of indazole N1- and N2-(beta-D-ribonucleosides). AB - The regioselective synthesis of 4-nitroindazole N1- and N2-(beta-D ribonucleosides) (8, 9, 1b and 2b) is described. The N1-regioisomers are formed under thermodynamic control of the glycosylation reaction [fusion reaction or Silyl Hilbert-Johnson glycosylation for 48 h (66%)], while the kinetic control (Silyl Hilbert-Johnson glycosylation for 5 h) afforded only the N2-isomer (64%). The structures of the nucleosides 1b and 2b were assigned by single crystal X-ray analyses. The 4-amino-N1-(beta-D-ribofuranosyl)-1H-indazole (3b) was obtained from the nitro nucleoside 1b by catalytic hydrogenation. Compound 3b shows fluorescence while the 4-nitroindazole nucleosides 1b and 2b do not possess this property. PMID- 15043150 TI - Synthesis of tunicaminyluracil derivatives. AB - A tunicaminyluracil derivative, which is a key component of the tunicamycin nucleoside antibiotics, was synthesized using a samarium diiodide (SmI2) mediated aldol reaction and intramolecular Pummerer reaction as the key steps. The alpha phenylthio ketone 11, the precursor of the samarium enolate, was prepared from D galactose. Treatment of 11 with SmI2 at -40 degrees C resulted in complete conversion to the corresponding samarium enolate, and subsequent addition of uridine 5'-aldehyde 12 afforded the desired aldol products 13a,b. Compound 13a was converted to the sulfoxide 15 by a sequential diastereoselective reduction of the ketone and an oxidation with mCPBA. Activation of 15 with Tf2O provided the desired cyclized compound 17. In this reaction, the aldol product 13a was also obtained as a consequence of a competitive intramolecular version of DMSO oxidation via a 7-membered ring intermediate. Compound 18 or 19 are ready for use as a glycosyl donor in glycosylations to provide a range of analogues as potential glycosyltransferase inhibitors as well as related natural products. PMID- 15043151 TI - New sequential-assignment routes of nucleic acid NMR signals using a [5'-(13)C] labeled DNA dodecamer. AB - NMR signal assignments for DNA oligomers have been performed by the well established sequential assignment procedures based on NOESY and COSY. The H4'/H5'/H5'' resonance region is congested and difficult to analyze without the use of isotope-labeled DNA oligomers. Here a DNA dodecamer constructed with 2' deoxy[5'-(13)C]ribonucleotides, 5'-d(*C*G*C*G*A*A*T*T*C*G*CG)-3' (*N = [5' (13)C]Nucleotide), was prepared in an effort to analyze the H4'/H5'/H5'' resonance region by 2D 1H-13C HMQC-NOESY. In the C5' and H1' resonance region, weak and strong cross peaks for C5'(i)-H1'(i) and C5'(i)-H1'(i-1), respectively, were found, thus enabling the sequential assignment within this region. A similar sequential assignment route was found between C5' and H2''. Proton pair distances evaluated from the canonical B-DNA as well as A-DNA indicated that these sequential-assignment routes on a 2D 1H-13C HMQC-NOESY spectrum work for most nucleic acid stem regions. PMID- 15043152 TI - Inhibitors of adenosine deaminase: continued studies of structure-activity relationships in analogues of coformycin. AB - Synthesis and adenosine deaminase (ADA) inhibitory activity of two analogues of coformycin, containing the imidazo[4,5-e][1,2,4]triazepine ring system, have been reported as part of the structure-activity relationship (SAR) studies to explore the factors responsible for the extremely tight-binding characteristics of coformycins to ADA. PMID- 15043153 TI - Use of both direct and indirect 13C tags for probing nitrogen interactions in hairpin ribozyme models by 15N NMR. AB - We have used the synthesis and 15N NMR study of separate loop A and loop B domains of the hairpin ribozyme to demonstrate that multiple 15N atoms can be incorporated into an RNA strand and be unambiguously distinguished through a combination of direct and indirect tagging by 13C atoms. Absence of 15N chemical shift changes shows that the G8N1 in loop A does not become deprotonated up to pH 8, and that the G21N7 of loop B does not bind to Mg2+. PMID- 15043154 TI - The interaction of per-O-acetylated acyclic 1-(1-butylindol-3-yl)-1-deoxy-ketoses with silylated uracil. AB - The per-O-acetylated open chain derivatives of 1-(1-butylindol-3-yl)-1-deoxy-1-L sorbose and 1-(1-butylindol-3-yl)-1-deoxy-L-tagatose, which are readily available by alkaline degradation of 1-butylascorbigen followed by acetylation, were used in a nucleoside-type synthesis. The interaction of these ketoses derivatives with bis-(trimethylsilyl)-uracil yielded in each case a mixture of (E)-2,4,5,6-tetra-O acetyl-1-(1-butylindol-3-yl)-1,3-dideoxy-3-(uracil-1-yl)-L-xylo-hexa-1-enitol and (E)-2,4,5,6-tetra-O-acetyl-1-(1-butylindol-3-yl)-1,3-dideoxy-3-(uracil-1-yl)-L lyxo-hexa-1-enitol, which were separated by preparative HPLC. The deacetylation of each of these compounds by MeONa in MeOH produced a mixture of 1-(1-butylindol 3-yl)-1,3-dideoxy-4-O-methyl-3-(uracil-1-yl)-alpha-L-sorbopyranose and 1-(1 butylindol-3-yl)-1,3-dideoxy-4-O-methyl-3-(uracil-1-yl)-beta-D-fructopyranose, which were also separated by HPLC, the structures were confirmed by NMR. PMID- 15043155 TI - Tissue disposition of 5-o-carboranyluracil--a novel agent for the boron neutron capture therapy of prostate cancer. AB - The carboranyl nucleotides beta-D-5-o-carboranyl-2'-deoxyuridine (D-CDU), 1-(beta L-arabinosyl)-5-o-carboranyluracil (D-ribo-CU) and the nucleotide base 5-o carboranyluracil (CU), were developed as sensitizers for boron neutron capture therapy (BNCT). A structure activity study was initiated to determine the agent most suitable for targeting prostate tumors. Cellular accumulation studies were performed using LNCaP human prostate tumor cells, and the respective tumor disposition profiles were investigated in male nude mice bearing LNCaP and 9479 human prostate tumor xenografts in their flanks. D-CDU achieved high cellular concentrations in LNCaP cells and up to 2.5% of the total cellular compound was recovered in the 5'-monophosphorylated form. In vivo concentrations of D-CDU were similar in LNCaP and 9479 tumor xenografts. Studies in 9479 xenografted bearing mice indicated that increasing the number of hydroxyl groups in the sugar moeity of the carboranyl nucleosides corresponded with an increased rate and extent of renal elimination, shorter serum half-lives and an increased tissue specificity. Tumor/normal prostate ratios were greatest with the nucleoside base CU. These studies indicate that similar nucleoside analogues and bases may have different tissue affinities and retention properties, which should be considered when selecting agents for sensitizing specific tumors for eventual BNCT treatment. CU was found to be the most suitable compound for further development to treat prostate cancer. PMID- 15043156 TI - Synthesis of 4-formyl-4-imidazolin-2-one nucleosides, isomers of uridine and 2' deoxyuridine. AB - The syntheses of the ribo- and deoxyribonucleoside derivatives of 4-formyl-4 imidazolin-2-one, isosteric isomers of uridine and 2'-deoxyuridine, respectively, were carried out by ring contraction of the corresponding 5-bromouracil nucleosides, followed by conversion of the carboxyl side-chain of the products to the respective carboxaldehyde derivatives. PMID- 15043157 TI - Synthesis of azole nucleoside 5'-monophosphate mimics (P1Ms) and their inhibitory properties of IMP dehydrogenases. AB - IMPDH inhibitors have potential antimicrobial, anticancer and immunomodulatory effects. Nucleoside inhibitors of IMPDH exert their inhibitory effects via nucleoside 5'-MPs. Conversion of nucleoside analogs to NMPs by cellular nucleoside kinases is not assured, and usually is inefficient. In order to bypass cellular phosphorylation, a series of azole nucleoside 5'-MP mimics (P1Ms) based on ribavirin, EICAR and bredinin were synthesized and screened against human and C. albicans IMP dehydrogenises. P1Ms 8, 16, 25, 28 and 29 demonstrated substantial IMPDH inhibition with Ki values in low micromolar range. PMID- 15043158 TI - Effect of imino group of a linker arm at the C5 position of a pyrimidine nucleoside on the thermal stabilities of DNA/DNA and DNA/RNA duplexes. AB - The modified ODN's bearing C5-substituted 2'-deoxyuridine derivative were synthesized by a post-synthetic modification with an unsymmetrical triamine. The effect of the C5-substituent on the duplex formation with complementary DNA or RNA differed with the position of an imino group in the linker-arms. PMID- 15043159 TI - Nucleosides. IX. Synthesis of purine N(3),5'-cyclonucleosides and N(3),5'-cyclo 2',3'-seconucleosides via Mitsunobu reaction as TIBO-like derivatives. AB - The Mitsunobu reaction was applied to prepare, in one step, purine N(3),5' cyclonucleosides 10a-d. A subsequent ring opening in the ribose moiety of the resultant N(3),5'-nucleosides by sodium periodate led to the corresponding N(3),5'-cyclo-2',3'-seconucleosides. These products consist of 5-, 6-, and 7 membered tricyclic system which is the basic skeleton of TIBO derivatives, known antiviral agents. PMID- 15043160 TI - Uridine recognition motifs of human equilibrative nucleoside transporters 1 and 2 produced in Saccharomyces cerevisiae. AB - The sugar moiety of nucleosides has been shown to play a major role in permeant transporter interaction with human equilibrative nucleoside transporters 1 and 2 (hENT1 and hENT2). To better understand the structural requirements for interactions with hENT1 and hENT2, a series of uridine analogs with sugar modifications were subjected to an assay that tested their abilities to inhibit [3H]uridine transport mediated by recombinant hENT1 and hENT2 produced in Saccharomyces cerevisiae. hENT1 displayed higher affinity for uridine than hENT2. Both transporters barely tolerated modifications or inversion of configuration at C(3'). The C(2')-OH at uridine was a structural determinant for uridine-hENT1, but not for uridine-hENT2, interactions. Both transporters were sensitive to modifications at C(5') and hENT2 displayed more tolerance to removal of C(5')-OH than hENT1; addition of an O-methyl group at C(5') greatly reduced interaction with either hENT1 or hENT2. The changes in binding energies between transporter proteins and the different uridine analogs suggested that hENT1 formed strong interactions with C(3')-OH and moderate interactions with C(2')-OH and C(5')-OH of uridine, whereas hENT2 formed strong interactions with C(3')-OH, weak interactions with C(5')-OH, and no interaction with C(2')-OH. PMID- 15043161 TI - Anti-cowpox virus activities of certain adenosine analogs, arabinofuranosyl nucleosides, and 2'-fluoro-arabinofuranosyl nucleosides. AB - Nucleoside analogs were investigated for their potential to inhibit cowpox virus (a surrogate for variola and monkeypox viruses) in cell culture and in lethal respiratory infections in mice. Cell culture antiviral activity was determined by plaque reduction assays, with cytotoxicity determined by cell proliferation assays. Selectivity indices (SI's, 50% cytotoxic concentration divided by 50% virus-inhibitory concentration) were determined for 15 compounds. Three arabinofuranosyl (Ara) nucleosides showed activity in mouse mammary tumor (C127I) cells: guanine (Ara-G), thymine (Ara-T), and adenine (Ara-A) with SI's of 113, 61, and 95, respectively. The 2'-fluoro-Ara nucleosides of 5-F-cytosine (FIAC), 5 methyluracil (FMAU), and 5-iodouracil (FIAU) exhibited SI's of 148, 77, and 29, respectively. Other potent compounds included cidofovir (a positive control) and 3'-O-methyladenosine, with SI values of 164 and 56, respectively. In general, assays performed in African green monkey kidney (Vero) cells produced lower SI's than in C127I cells, except for 5-iodo-2'-deoxyuridine (IDU) which had an SI of > 71 in Vero cells and 3.1 in C127I cells. Intranasal infection of mice with cowpox virus was followed a day later by twice daily intraperitoneal treatment with compounds for 5 days. Ara-A was active at 300 mg/kg/day (40% survival), FMAU at 100 mg/kg/day (70% survival), and cidofovir (given for 1 day only) at 100 mg/kg (80-100% survival). None of the other compounds, including IDU, prevented death nor delayed the time to death. Cidofovir had the best potential for treating orthopoxvirus infections of those tested. PMID- 15043162 TI - Synthesis and evaluation of a novel synthetic phosphocholine lipid-AZT conjugate that double-targets wild-type and drug resistant variants of HIV. AB - INK-20, a synthetic phosphocholine lipid-AZT conjugate, was evaluated for antiviral activity against wild-type HIV-1, a matched pair of pre-AZT and post AZT and multidrug resistant clinical isolates. In addition, it was tested for activity against viruses resistant to nucleoside (AZT, 3TC) and nonnucleoside (nevirapine) reverse transcriptase and protease (saquinavir) inhibitors using the syncytial plaque reduction assay for infectious virus multiplication. The EC50 values were 0.004, and 0.005 microM against wild-type HIV-1 for INK-20 and AZT, respectively. INK-20 showed little or no cytotoxicity when assayed in CEM-SS cells and four other cell types including PBMC. This resulted in a selective index of > 25,000 and > 20,000 for INK-20 and AZT, respectively. When tested against a matched pair of pre-AZT and post-AZT clinical isolates, the EC50 values were 0.01 and 0.03 microM for INK-20 and 0.0005 and 0.33 microM for AZT, respectively. INK-20 had moderate to good activity against two other AZT resistant variants and very good activity against a multi-drug resistant clinical isolate compared to marked resistance of these viruses to AZT alone. INK-20 retained significant activity against viruses resistant to 3TC, nevirapine, and saquinavir. The synthetic phosphocholine lipid-AZT conjugate INK-20 represents a novel class of anti-HIV compounds, which may provide new strategies for the treatment of HIV drug-resistant variants. PMID- 15043163 TI - Enhanced DNA-directed effects of FdUMP[10] compared to 5FU. AB - FdUMP[N] molecules and conjugates are much more effective at inhibiting the proliferation of human tumor cells than is the widely used anticancer drug 5 fluorouracil (5FU). We have evaluated the inhibition of thymidylate synthase (TS), the extent of DNA damage, cell cycle arrest, and the induction of apoptosis by FdUMP[10] and 5FU in the human colorectal cancer cell line HT29. The magnitude and duration of TS inhibition following exposure of HT29 cells to FdUMP[10] at 1 x 10(-8) M was greater than that which occurred following exposure of these cells to 5FU at 1 x 10(-6) M. FdUMP[10] exposure also resulted in much more extensive DNA damage to HT29 cells than occurred following exposure to 100-fold higher concentrations of 5FU. Although exposure of HT29 cells to both drugs resulted in S-phase arrest, more complete accumulation of cells in S-phase was achieved following FdUMP[10] exposure at much lower drug concentrations. FdUMP[10] was also much more effective at inducing apoptosis in HT29 cells than was 5FU. The results are consistent with FdUMP[10] being much more efficient that 5FU at inducing DNA damage that results in apoptotic cell death in colon cancer cells. PMID- 15043164 TI - Synthesis of 2'-substituted MMI linked nucleosidic dimers: an optimization study in search of high affinity oligonucleotides for use in antisense constructs. AB - The synthesis of a series of methylene(methylimino) (MMI) linked oligodeoxyribonucleotide dimers modified at the 2'-position with fluoro and/or methoxy groups and their incorporation into different sequences has been accomplished. From these dimers, bis 2'-OMe MMI dimer was selected for further studies based on its synthetic accessibility and the increased thermodynamic stability conferred upon oligonucleotides incorporating this modification. PMID- 15043165 TI - Synthesis of D-altritol nucleosides with a 3'-O-tert-butyldimethylsilyl protecting group. AB - Four D-altritol nucleosides with a 3'-O-tert-butyldimethylsilyl protecting group are synthesized (base moieties are adenine, guanine, thymine and 5 methylcytosine). The nucleosides are obtained by ring opening reaction of 1,5:2,3 dianhydro-4,6-O-benzylidene-D-allitol. Optimal reaction circumstances (NaH, LiH, DBU, phase transfer, microwave irridation) for the introduction of the heterocycles are base-specific. For the introduction of the 3'-O-silyl protecting group, long reaction times and several equivalents of tert-butyldimethylsilyl chloride are needed. PMID- 15043166 TI - Nucleoside analogues exerting antiviral activity through a non-nucleoside mechanism. AB - In analogy with maribavir [1-(beta-L-ribofuranosyl)-isopropylamino-5,6 dichlorobenzimidazole], a nucleoside analogue that acts against human cytomegalovirus (HCMV) by a non-nucleoside mechanism, here I present three other examples of classes of nucleoside analogues (i.e. bicyclic furo[2,3-d]pyrimidine as well as HEPT and TSAO derivatives) that act against either HCMV or human immunodeficiency virus (HIV) through a non-nucleoside mode of action. PMID- 15043167 TI - A2B adenosine receptor agonists: synthesis and biological evaluation of 2 phenylhydroxypropynyl adenosine and NECA derivatives. AB - In the search for agonists for the elusive A2B adenosine receptor subtypes, 2 phenylhydroxypropynyl-5'-N-methylcarboxamido adenosine (PHPMECA, 14), 2 phenylhydroxypropynyl-5'-N-propylcarboxamido adenosine (PHPPECA, 15), and N6 ethyl-2-phenylhydroxypropynyl-5'-N-ethylcarboxamidoadenosine (19) were synthesized on the basis that introduction of alkynyl chains in 2-position of adenosine derivatives resulted in reasonably good A2B potency compared to NECA [see N6-ethyl-2-phenylhydroxypropynyl adenosine (5) EC50 = 1,700 nM and 2 phenylhydroxypropynyl-5'-N-ethylcarboxamido adenosine (PHPNECA, 8) EC50 = 1,100 nM, respectively]. Radioligand binding studies and adenylyl cyclase assays, performed with recently cloned human A1, A2A, A2B, and A3 adenosine receptors, showed that these modifications produced a decrease in potency at A2B receptor, as well as a general reduction in affinity at the other receptor subtypes. On the other hand, the contemporary presence of an ethyl substituent in N6-position and of a 4'-ethylcarboxamido group in the same compounds led to (R,S)-N6-ethyl-2 phenylhydroxypropynyl-5'-N-ethylcarboxamidoadenosine and (S)-N6-ethyl-2 phenylhydroxypropynyl-5'-N-ethylcarboxamidoadenosine, which did not show the expected increase in potency at A2B subtype. Hence, (S)-2-phenylhydroxypropynyl 5'-N-ethylcarboxamidoadenosine [(S)-PHPNECA] with EC50 A2B = 220 nM remains the most potent agonist at A2B receptor reported so far. PMID- 15043168 TI - Monitoring the intracellular metabolism of nucleoside phosphoramidate pronucleotides by 31P NMR. AB - The intracellular metabolism of 3'-azido-3'-deoxythymidine (AZT)-(L)-tryptophan methyl ester phosphoramidate (L-ATO) and AZT-(L)-phenylalanine methyl ester phosphoramidate (L-APO) by the human T-lymphoblastoid cell line CCRF-CEM (CEM 1.3) and peripheral blood mononuclear cell line (PBMC) was investigated with high field 31P NMR spectroscopy. The AZT amino acid phosphoramidates were shown to accumulate intracellularly and to be readily converted into AZT-MP by both tissues types. Thus, the efficient delivery of nucleoside monophosphates to cells can be facilitated by nucleoside phosphoramidate pronucleotides. PMID- 15043169 TI - Synthesis of nucleoside libraries on solid support. I. N2,N6-disubstituted diaminopurine nucleosides. AB - Starting with 2-iodo-6-chloro-9-(beta-D-ribofuranosyl)purine, a library of more than 1,300 N2,N6-polysubstituted diaminopurine nucleosides was created. The starting material was condensed with a polystyrene monomethoxytrityl resin and a pool of primary and secondary amines was used to displace the 6-chloro atom with high regioselectivity. The 2-iodo was subsequently displaced by various primary amines. Nucleosides were cleaved from the resin with hexafluoroisopropanol solutions. A majority of compounds reached a purity of more than 80% without the need for any type of purification. PMID- 15043170 TI - Synthesis of nucleoside libraries on solid support. II. 2,6,8-Trisubstituted purine nucleosides using 8-bromoguanosine as precursor. AB - A series of 2,6,8-trisubstituted purine nucleoside libraries was prepared by parallel solid-phase synthesis using 8-bromoguanosine as a common synthetic precursor. Polystyrene-methoxytrityl chloride resin was linked to the N2 or O5' position of the guanosine analogues. 8-Bromoguanosine was derivatized at the C8 position via carbon-carbon bond formation. Nucleophilic aromatic substitution at C2 and/or C6 positions with various amines produced two series of purine nucleoside libraries with very diverse substitution. PMID- 15043171 TI - Oligonucleotide conjugates of Nile Red. AB - Oligonucleotides containing long chain amine on uridine-2'-carbamate have been covalently modified with Nile Red, a hydrophobic long wavelength fluorescent benzophenoxazine dye. The fluorescence of the dye is quenched in oligonucleotide conjugates. Thermal denaturation studies show considerable interactions of Nile Red with DNA-DNA duplexes. PMID- 15043172 TI - Conformational flexibility in a triazole nucleoside derivative: 4-cyano-5 cyanomethyl-1-(2,3,5-tri-O-acetyl-beta-D-ribofuranosyl)-1,2,3-triazole. AB - The crystal-structure determination of the molecular structure of the hydrophobic compound, 4-cyano-5-cyanomethyl-1-(2,3,5-tri-O-acetyl-beta-D-ribofuranosyl)-1,2,3 triazole, C16H17N5O7, provides us with two different conformations of ribofuranosyl moieties [(C2'-exo, C3'-endo) and C2'-exo] with two markedly different N-glycosidic angles. There are two molecules in the asymmetric unit of the crystal and the overall stereochemistry of the molecules are influenced predominantly by weak intramolecular bifurcated and trifurcated hydrogen bonds of the type C-H...O and C-H...N, where endo-H atoms attached to C2' and C3' are involved. The molecules in the crystal are interconnected with longer intermolecular bonds of the same type. There are empty channels (occupying 14.0% of the whole volume of the unit cell), which are extended along b-axis of the entire crystal. PMID- 15043173 TI - Chronic illness, comorbidities, and the need for medical generalism. PMID- 15043174 TI - Comorbidity: implications for the importance of primary care in 'case' management. AB - BACKGROUND: Although comorbidity is very common in the population, little is known about the types of health service that are used by people with comorbid conditions. METHODS: Data from claims on the nonelderly were classified by diagnosis and extent of comorbidity, using a case-mix measure known as the Johns Hopkins Adjusted Clinical Groups, to study variation in extent of comorbidity and resource utilization. Visits of patients (adults and children) with 11 conditions were classified as to whether they were to primary care physicians or to other specialists, and whether they involved the chosen condition or other conditions. RESULTS: Comorbidity varied within each diagnosis; resource use depended on the degree of comorbidity rather than the diagnosis. When stratified by degree of comorbidity, the number of visits for comorbid conditions exceeded the number of visits for the index condition in almost all comorbidity groups and for visits to both primary care physicians and to specialists. The number of visits to primary care physicians for both the index condition and for comorbid conditions almost invariably exceeded the number of visits to specialists. These patterns differed only for uncommon conditions in which specialists played a greater role in the care of the condition, but not for comorbid conditions. CONCLUSIONS: In view of the high degree of comorbidity, even in a nonelderly population, single-disease management does not appear promising as a strategy to care for patients. In contrast, the burden is on primary care physicians to provide the majority of care, not only for the target condition but for other conditions. Thus, management in the context of ongoing primary care and oriented more toward patients' overall health care needs appears to be a more promising strategy than care oriented to individual diseases. New paradigms of care that acknowledge actual patterns of comorbidities as well as the need for close coordination between generalists and specialists require support. PMID- 15043175 TI - Descriptions of barriers to self-care by persons with comorbid chronic diseases. AB - BACKGROUND: Chronic medical conditions often occur in combination, as comorbidities, rather than as isolated conditions. Successful management of chronic conditions depends on adequate self-care. However, little is known about the self-care strategies of patients with comorbid chronic conditions. OBJECTIVE: Our objective was to identify perceived barriers to self-care among patients with comorbid chronic diseases. METHODS: We conducted semistructured personal interviews with 16 adults from 4 urban family practices in the CaReNet practice based research network who self-reported the presence of 2 or more common chronic medical conditions. Using a free-listing technique, participants were asked, "Please list everything you can think of that affects your ability to care for your medical conditions." Responses were analyzed for potential barriers to self care. RESULTS: Participants' responses revealed barriers to self-care, including physical limitations, lack of knowledge, financial constraints, logistics of obtaining care, a need for social and emotional support, aggravation of one condition by symptoms of or treatment of another, multiple problems with medications, and overwhelming effects of dominant individual conditions. Many of these barriers were directly related to having comorbidities. CONCLUSIONS: Persons with comorbid chronic diseases experience a wide range of barriers to self-care, including several that are specifically related to having multiple medical conditions. Self-management interventions may need to address interactions between chronic conditions as well as skills necessary to care for individual diseases. PMID- 15043176 TI - Patient education for informed decision making about prostate cancer screening: a randomized controlled trial with 1-year follow-up. AB - PURPOSE: The efficacy of prostate cancer screening is uncertain, and professional organizations recommend educating patients about potential harms and benefits. We evaluated the effect of a videotape decision aid on promoting informed decision making about prostate cancer screening among primary care patients. METHODS: A group of 160 men, 45 to 70 years of age, with no history of prostate cancer, were randomized to view or not to view a 20-minute educational videotape before a routine office visit at a university-based family medicine clinic. The subjects were contacted again 1 year after their visit to assess their receipt of prostate cancer screening (digital rectal examination [DRE] or prostate-specific antigen [PSA] testing), their satisfaction with their screening decision, and knowledge retention since the baseline assessment. RESULTS: Follow-up assessments were completed for 87.5% of the intervention subjects and 83.8% of the control subjects. The rate of DRE did not differ between the 2 groups. Prostate-specific antigen testing was reported by 24 of 70 (34.3%) intervention subjects and 37 of 67 (55.2%) control subjects (P = .01). African American men were more likely to have had PSA testing (9 of 16, 56.3%) than were white men (13 of 46, 28.3%) (P = .044). Satisfaction with the screening decision did not differ between the study groups. Intervention subjects were more knowledgeable of prostate cancer screening than were control subjects, although these differences declined within 1 year (P < .001). CONCLUSIONS: Decision aids for prostate cancer screening can have a long-term effect on screening behavior and appear to promote informed decision making. PMID- 15043177 TI - Glycemic control in medical inpatients with type 2 diabetes mellitus receiving sliding scale insulin regimens versus routine diabetes medications: a multicenter randomized controlled trial. AB - PURPOSE: Hospitalized patients with type 2 diabetes mellitus traditionally receive insulin on a sliding-scale regimen, but the benefits of this approach are unclear. The purpose of this study was to compare the effects of the sliding scale insulin regimen with those of routine diabetes medications on hyperglycemia, hypoglycemia and length of hospitalization in diabetic patients hospitalized for other conditions. METHODS: This was a multicenter, randomized controlled trial conducted in family medicine inpatient services. One hundred fifty-three patients with type 2 diabetes mellitus hospitalized for other conditions were randomized to receive routine diabetes medications (control) or the combination of a standard sliding-scale insulin regimen and routine diabetes medications (intervention). The outcome measures included frequency of hyperglycemia and hypoglycemia (glycemic events), and length of hospitalization. RESULTS: No differences were identified between treatment groups in the frequency of glycemic events. In the intervention group, 33.3% of patients developed hyperglycemia compared to 34.6% in the control group (P = .87). Six patients developed hypoglycemia in the intervention group, compared with 7 in the control group (P = .83). There was no difference in length of hospitalization (P = .86). Regardless of treatment assignment, patients receiving intermediate-acting insulin (OR, 2.8; 95% CI, 1.2-6.5), those with blood glucose values greater than 250 mg/dL at baseline (OR, 6.3; 95% CI, 2.3 - 17.2) and those receiving corticosteroids (OR, 9.1; 95% CI, 3.1 - 27.0) were more likely to have glycemic events. CONCLUSIONS: The use of the sliding scale insulin regimen in combination with routine diabetes medications does not affect the rate of hyperglycemia, hypoglycemia or length of hospitalization in patients with type 2 diabetes mellitus hospitalized for other conditions. PMID- 15043178 TI - A Native American community with a 7% cesarean delivery rate: does case mix, ethnicity, or labor management explain the low rate? AB - PURPOSE: Cesarean delivery rates vary widely across populations. Studying communities with low rates of cesarean delivery may identify practices that can lower the cesarean rate. METHODS: A population-based historical cohort study included all pregnant women (N = 1132) from 1992 through 1996 in a predominantly Native American region of northwestern New Mexico known to have a high prevalence of gestational diabetes and preeclampsia. The outcomes studied included delivery type (eg, cesarean, operative vaginal, spontaneous vaginal), indication for cesarean delivery, presence of obstetrical risk factors, and use of labor induction or augmentation. RESULTS: The cesarean delivery rate of the study group (7.3%) was only 35% of the 1996 US rate of 20.7%. Among study participants, the relative risk of a primary cesarean delivery for dystocia was 0.22 (95% CI, 0.14, 0.35). Trial of labor after cesarean delivery was attempted by 93% of study participants compared with 42% of women nationwide in 1994. The cesarean delivery rates for women with diabetes in pregnancy (11.5% versus 35.4%) and preeclampsia (14.8% versus 37.4%) were significantly lower than nationwide rates. Case-mix analysis comparison with a standardized population and comparison of standard (ie, term, singleton, vertex) primiparous women demonstrate that the low rate of cesarean delivery was not because of a lower prevalence of risk factors. CONCLUSIONS: The community's low rate of cesarean delivery is primarily the result of a decreased use of cesarean delivery for labor dystocia and an almost universal acceptance of trial of labor after cesarean delivery. Cultural attitudes toward childbirth, design of the perinatal system, and genetic factors also may explain the low rate of cesarean delivery. PMID- 15043181 TI - Making care safe. PMID- 15043180 TI - The face of cancer. AB - Two voices tell the story of an evolving doctor-patient relationship. Seizures are the first indication of squamous cell lung cancer metastatic to the brain. Both doctor and patient describe the course of the illness, treatment, and remarkable recovery, each from his own perspective. The patient describes how the cancer became a creative as well as a destructive force, expressed in his painting and new approach to life. The essay concludes by discussing limitations of the biomedical model of disease in this case and the complex two-way nature of the healing relationship between doctor and patient. PMID- 15043179 TI - Applying a risk-adjustment framework to primary care: can we improve on existing measures? AB - Outcome-based performance measurement and prospective payment are common features of the current managed care environment. Increasingly, primary care clinicians and health care organizations are being asked to assume financial risk for enrolled patients based on negotiated capitation rates. Therefore, the need for methods to account for differences in risk among patients enrolled in primary care organizations has become critical. Although current risk-adjustment measures represent significant advances in the measurement of morbidity in primary care populations, they may not adequately capture all the dimensions of patient risk relevant to primary care. We propose a risk-adjustment framework for primary care that incorporates clinical features related to patients' health status and nonclinical factors related to patients' health behaviors, psychosocial factors, and social environment. Without this broad perspective, clinicians with more unhealthy and more challenging populations are at risk of being inadequately compensated and inequitably compared with peers. The risk-adjustment framework should also be of use to health care organizations that have been mandated to deliver high-quality primary care but are lacking the necessary tools. PMID- 15043182 TI - Refining the paradigm: the transition from recertification to maintenance of certification. PMID- 15043183 TI - Research and the Society of Teachers of Family Medicine. PMID- 15043184 TI - 'Swotting' departments: issues and challenges in academic family medicine. PMID- 15043185 TI - Health Services Researcher Lu Ann Aday: primary care and health services researchers as partners. PMID- 15043186 TI - The basic biology of metastasis. PMID- 15043187 TI - Clinical aspects of bone metastases in prostate cancer. PMID- 15043188 TI - Animal models of bone metastasis. AB - Animal models will continue to be indispensable to investigate the pathogenesis of bone metastasis in vivo, conduct preclinical chemotherapeutic, chemoprevention and genetic therapy studies, test gene delivery mechanisms, and identify metastasis suppressor and inducer genes. It is likely that the bone marrow microenvironment, such as the endothelial cells, stromal cells, hematopoietic cells, bone cells, and the intercellular matrix play important roles in the localization and clonal growth of cancer cells in bone. Given the complexity of bone metastasis, many genes are expected to be involved in the pathogenesis and few are likely indispensable. The use of genomic and proteomic approaches to study these animal models will identify key targets for therapeutic intervention. As we further refine these models and use imaging for real-time evaluation of cells, and eventually target genes, these models will more closely mirror human disease and will hopefully become more predictive of the human response to therapy. PMID- 15043189 TI - MIP-1 alpha and myeloma bone disease. AB - Figure 5 is a proposed model for MIP-1alpha's effects on myeloma bone disease. MIP-1alpha is produced by myeloma cells and directly stimulates OCL formation. In addition MIP-1alpha enhances adhesive interactions between myeloma cells and marrow stromal cells increasing expression of RANKL and IL-6, which further increase bone destruction and tumor burden. The recent evidence from our group and others lead to the conclusion that MIP-1alpha is an important mediator in the debilitating bone destruction in multiple myeloma. Blocking MIP-1alpha expression may have profound effects on myeloma cell growth, homing, and bone destruction in this in vivo model of myeloma. These data suggest that antagonists that decrease MIP-1alpha activity in vivo or blocking MIP-1alpha signaling by neutralizing its receptor may provide therapeutic alternatives for treating patients with myeloma to decrease both their tumor burden and bone destruction. PMID- 15043190 TI - Type I collagen-mediated changes in gene expression and function of prostate cancer cells. AB - In this study, cDNA microarrays were used to characterize gene expression changes elicited in prostate cancer cells by plating them on type I collagen. The results clearly reveal changes in the expression of genes associated with cellular signaling, cellular metabolism, gene transcription and gene translation which are indicative of cells that are actively proliferating. Together these results suggest that these changes in the gene expression profiles mediated by type I collagen may influence the proliferative capacity of prostate cancer cells in the bone microenvironment and facilitate development of prostate cancer bone metastases. Additionally, the microarray approach provides an invaluable tool to determine and track changes in gene expression in numerous disease states including prostate cancer. This technology is certain to facilitate discovery of new therapeutic gene targets. PMID- 15043191 TI - PTHrP and skeletal metatasis. PMID- 15043192 TI - Opg, RANKl, and RANK in cancer metastasis: expression and regulation. PMID- 15043193 TI - Matrix metalloproteinaes and bone metastasis. PMID- 15043194 TI - Endothelins in bone cancer metastases. AB - Most evidence indicates that osteoblastic bone metastases are due to tumor produced factors that stimulate the osteoblast. This review supports a causal role for ET-1. Based on our results, we propose a model to explain the tumor cell and bone interactions that are responsible for the osteoblastic response (Figure 2). Tumor cells housed in bone produce factors, such as ET-1, stimulate osteoblast activity. This results in the abundant and disorganized new bone formation that is characteristic of osteoblastic metastases. The effects of ET-1 to stimulate bone formation are mediated by ETA receptors on the osteoblast. ETA receptor inhibition successfully blocked osteoblastic bone metastases in a mouse model. These receptor antagonists are currently in clinical trials for advanced prostate cancer and bone metastases (Stephenson, 2001; Carducci et al., 2002; 2003). Therefore, the molecular mechanisms responsible for osteoblastic metastases are complex and involve bi-directional interactions between tumor cells and bone. Elucidation of the interactions at a molecular level can identify therapeutic targets for osteoblastic metastases. Although ET-1 and ETA receptors are potential targets for this devastating complication of cancer (Remuzzi et al., 2003), they are certainly not the only ones. The rapid pace of metastasis research, will not only expand our therapeutic armamentarium against bone metastases, but will also provide insight into achieving the ultimate goal: the prevention of cancer metastases to bone. PMID- 15043195 TI - Bisphosphonate actions on bone and visceral metastases. PMID- 15043196 TI - Gene therapy for prostate cancer bone metastasis. Gene therapy targeting bone metastasis. PMID- 15043197 TI - Cancer cells homing to bone: the significance of chemotaxis and cell adhesion. PMID- 15043198 TI - Histological, immunophenotypic and histomorphometric characterization of prostate cancer bone metastases. PMID- 15043199 TI - Pharmacological cyclin-dependent kinase inhibitors as HIV-1 antiviral therapeutics. AB - Human immunodeficiency virus type 1 (HIV-1) can infect quiescent cells; however, viral production is restricted to actively proliferating cells. Recent evidence has indicated that HIV-1 viral proteins, Vpr and Tat, perturb the cell cycle to optimize HIV-1 replication. Vpr arrests the cell cycle at G2 by inactivating the cyclin B/cdk1 complex. Tat regulates the cell cycle by altering factors involved in proliferation and differentiation (i.e. the cdk inhibitor p21/waf1) and associating with cyclin/cdk complexes (i.e. cyclin E/cdk2, cyclin H/cdk7, and cyclin T/cdk9). These studies indicate the importance of host cellular factors, such as cyclin/cdk complexes, in regulating HIV-1 replication and therefore represent novel targets for antiviral therapeutics. Recently, the efficacy of pharmalogical cdk inhibitors (PCIs) in abrogating viral replication has been under development. To date there are 25-30 PCIs that have been synthesized against known cdks, several of which have been shown to inhibit HIV-1 and other AIDS-associated viruses in vitro and in vivo. Targeting these critical cyclin/cdk complexes needed for viral propagation may solve the problems inherent in current HAART therapy, including the emergence of drug-resistant viruses. Thus, PCIs have the potential to become novel therapeutic antiviral drugs that can inhibit HIV-1 transcription and opens the possibility of new avenues of treatment. PMID- 15043200 TI - Replicative senescence: the final stage of memory T cell differentiation? AB - One of the major obstacles to effective prolonged CD8 T cell control over HIV and other latent infections may be the intrinsic, genetically programmed barrier to unlimited proliferation that is characteristic of all normal human somatic cells. Replicative senescence, characterized extensively in cell culture for a variety of cell types, comprises both irreversible cell cycle arrest and striking changes in function. CD8 T cells with features similar to senescent CD8 T cell cultures (i.e., absence of CD28, inability to proliferate, telomeres in the 5-7 kb range, resistance to apoptosis) increase progressively during aging and in chronic HIV infection, suggesting that replicative senescence may be occurring in vivo, and, in fact, may constitute the final stage in the normal differentiation of human T cells. CD8 T cells with characteristics suggestive of senescence have also been implicated in modulating immune function and altering bone homeostasis. Further characterization of the underlying mechanism leading to the generation of senescent memory CD8 T cells and analysis of their functional attributes will help elucidate their role in HIV disease pathogenesis. PMID- 15043201 TI - The physiological relevance of CD4 receptor down-modulation during HIV infection. AB - Upon binding to the CD4 receptor the HIV envelope protein undergoes conformational changes that culminate in the fusion of the viral and cellular membranes. A few hours later, a sophisticated set of processes is initiated to ensure the down-modulation of the viral receptor. Three viral proteins participate in this process: Nef, Env, and Vpu, suggesting that this function is critical for virus replication. The mechanisms of action of these proteins have been extensively characterized. However, the physiological relevance of the virus induced CD4 down-modulation remains a focus of controversy, and the impact of this function on the viral life cycle has been underestimated. This review summarizes current hypotheses explaining why HIV needs to reduce expression of its own receptor, and discusses the experimental evidence supporting them. Recent findings indicate that efficient CD4 down-modulation is essential for the production of infectious particles, and highlight the importance of this function in HIV pathogenesis in vivo. Progression to disease correlates with enhanced viral induced CD4 down-modulation, and a subset of long-term nonprogressors carry viruses defective in this function. To date, the HIV-induced CD4 down-modulation has not been targeted for therapeutic intervention. Addressing the reasons why this function is so critical and understanding the interplay between viral and host factors governing surface expression of CD4 may provide clues for the development of new antiviral strategies. PMID- 15043202 TI - The impact of human allelic variation on HIV-1 disease. AB - Human allelic variants influence the susceptibility to HIV-1 infection and/or the subsequent rates of disease progression towards AIDS that average ten years, although they vary greatly among infected subjects. In this respect, studies involving multiply exposed persons who remain uninfected, long-term nonprogressors (who remain asymptomatic for fifteen years or more) or, in contrast, rapid progressors (who develop AIDS within two to three years post infection) as well as seroincident cohorts of patients with defined seroconversion dates have contributed to our comprehension of the effects of different natural human polymorphisms on HIV-1 disease. The current article aims at providing an up-to-date review on these polymorphisms that may be broadly classified into three general categories: (1) those that control viral entry into susceptible cells (namely, chemokine and chemokine receptor polymorphisms), (2) mutational variants of genes involved in immune regulation, such as interleukin 10 (IL-10), interleukin-4 (IL-4), tumor necrosis factor-alpha (TNF-alpha), and mannose-binding lectin (MBL), and (3) polymorphisms in genes involved in the adaptive immune recognition by T cells, [human leukocyte antigen (HLA) type]. Particular emphasis has been placed on the state-of-the-art biotechnological methodologies, such as "spectral genotyping" that utilizes molecular beacons in conjunction with polymerase chain reaction in real-time (real-time-PCR), which were developed to assist with the characterization of some of these determinants. Elucidating the functional role of these factors via the application of such biotechnological assays is expected to further enhance our understanding of the pathogenesis of HIV-1 infection, and, eventually, to enrich our therapeutic arsenal with novel antiviral agents or strategic approaches. PMID- 15043203 TI - Dendritic cells and the promise of therapeutic vaccines for human immunodeficiency virus (HIV)-1. AB - Treatment of human immunodeficiency virus (HIV)-1 infection with potent antiretroviral medications has provided considerable clinical benefit. However because of the limitations of current therapy, innovative approaches are needed to better control HIV-1 infection. Several studies have suggested that robust CD4+ T helper and CD8+ T cell responses may contribute to the immunologic control of HIV-1 infection in certain individuals. Most chronically infected patients, however, cannot control the infection and may benefit from stimulation of cellular immunity with immunotherapy. Dendritic cells (DCs) are potent professional antigen-presenting cells (APCs) and have a central role in directing the adaptive immune response to pathogens. The ability of DCs to stimulate naive T cells has long been thought to be crucial in initiating an effective immune response. As DCs are uniquely situated at the interface between the innate and adaptive immune systems, they are currently under intense scrutiny as potential adjuvants for vaccines in many clinical settings. Studies in healthy volunteers and patients with cancer have shown that antigen-pulsed DCs can boost both CD8+ and CD4+ T cell responses in vivo. Based on these promising findings, ex vivo antigen-pulsed DCs are being actively investigated in studies aimed at developing a therapeutic vaccine for individuals with HIV-1 infection. PMID- 15043204 TI - HIV-1 coreceptor usage, transmission, and disease progression. AB - HIV-1 coreceptor usage is believed to play a critical role in pathogenesis. To initiate infection, HIV-1 interacts with two cell surface receptors; CD4 is the primary receptor and the beta-chemokine receptors CCR5 and CXCR4 usually serve as secondary receptors. HIV-1 strains transmitted in vivo generally use CCR5. Viruses that use CCR5 (R5 viruses) appear to be associated with relatively stable infection. Years after chronic infection is established, CXCR4 utilizing strains emerge in approximately 50% of infected individuals. Viruses that use the coreceptor CXCR4 (X4 viruses) are associated with rapid CD4+ cell decline and disease progression. However, the mechanism by which X4 viruses are associated with accelerated disease progression has never been properly elucidated. For example, the association between X4 virus and acceleration of HIV-1 disease progression has been ascribed to the expanded spectrum of CXCR4+ precursor cells susceptible to infection by X4 strains. It has also been postulated that the decline of the host immune system associated with clinical AIDS may allow X4 viruses to evolve and replicate freely in late-stage infection. Discriminating between these and other alternatives is central to increasing our understanding of the fundamental pathogenic processes involved in HIV-1 infection. In this article, we critically review those studies published over the last few years that purport to examine the relationship between HIV-1 coreceptor usage, transmission, CD4+ T-cell depletion, and disease progression. PMID- 15043205 TI - Recombinant rhabdoviruses as potential vaccines for HIV-1 and other diseases. AB - The failure to develop vaccines to protect against important infectious diseases such as human immunodeficiency virus type I (HIV-1) or Hepatitis C virus (HCV) has increased the interest in new vaccine strategies. One of these methods is immunization with an attenuated recombinant viral vector expressing a foreign antigen, which could protect individuals from later exposure to the respective pathogen. A new method to recover a non-segmented negative-stranded RNA virus (NNSV) from cDNA was described for the first time for rabies virus (RV), a member of the rhabdovirus family. The same approach was successfully used for another rhabdovirus, vesicular stomatitis virus (VSV), and opened the possibility to use rhabdoviruses as vaccine vehicles and biomedical tools. Further research showed that the genomes of rhabdoviruses are highly flexible, easy to manipulate, and able to express large and even multiple foreign genes, and therefore are excellent vaccine candidates. In addition, it has been shown for both RV and VSV that their single surface glycoprotein G, which is responsible for attachment and fusion to the host cell, can functionally be replaced by other viral or cellular glycoproteins. This review gives an overview of the use of RV and VSV as promising new candidates in the fight against HIV-1 and other human diseases. PMID- 15043206 TI - The molecular epidemiology of HIV-1 in South and East Asia. AB - Foremost amongst human pathogens, the human immunodeficiency virus (HIV) exhibits a great genetic variability. The resultant fluidity of HIV enzymatic proteins allows them to remain functional whilst simultaneously evading immune surrveillance and antiretroviral therapy. This very variability, however, has been turned to powerful advantage in the study of the movement and evolution of HIV strains within and between human populations. Molecular analyses that estimate the relatedness between viral isolates, conducted in tandem with epidemiological studies, provide a new clarity of insight into the modes and routes of HIV transmission and epidemic spread. In this paper the principles underlying the molecular study of HIV and the achievements of this new field of epidemiology in southern and eastern Asia are reviewed. PMID- 15043207 TI - T cell immunity to HIV: defining parameters of protection. AB - In recent years, CD4 and CD8 T cell responses to HIV and SIV infection have been increasingly measured with the use of single-cell assays such as ELISPOT, MHC peptide oligomers, and cytokine flow cytometry. The results of these assays have been compared to those obtained with traditional bulk assays such as lymphoproliferation (by 3H-thymidine incorporation) and cytotoxicity (by 51Cr release). Such comparisons have led to some general understanding of the T cell responses that characterize progressive disease, long-term non-progressors, and individuals with viral suppression achieved by anti-retroviral therapy. In addition, prophylactic and therapeutic vaccine trials have also begun to use these assays of T cell immunity to gauge the immunogenicity of the vaccines. Whether such analyses will allow us to pick the best vaccine constructs, and whether they will provide us with an improved understanding of what constitutes protective cellular immunity to HIV, are major questions for the field. These questions will be examined in this review from the standpoint of current data and comparisons to other viral diseases. It is hypothesized that sophisticated multiparametric assays will be required to sort out the factors relevant for protective immunity in this complex disease. These parameters may include functional avidity, epitope breadth and specificity, proliferative capacity, cytokine repertoire, degree of anergy, and differentiation phenotype, as well as magnitude, of HIV-specific CD4 and CD8 T cells. PMID- 15043208 TI - HIV capsid assembly. AB - HIV Gag assembly is the first and most essential step in the formation of virus particles. Following protein synthesis, Gag relocates from ribosomes and forms a virus particle at the plasma membrane, using host factors and machinery. Early studies focused on mapping the regions within Gag required for assembly and identified three distinct domains (M, I, and L), although their precise locations within the three-dimensional structure of Gag awaited later study. In this review, I summarize the mapping results in the light of recent progress on Gag structures made by nuclear magnetic resonance and X-ray crystallography as well as further functional analysis. These data are largely consistent and provide sufficient information for an understanding of the interactions and functions of the assembly domains at a macromolecular level. Current studies have moved on to the identification of the host factors and machinery used in the process of Gag assembly. Cumulative data suggest that the dynamics of Gag assembly and transport are achieved not by simply using, but rather by taking control of, cellular machinery. Key area in the process include interactions with TSG101, L domain receptor which normally functions in the endosomal sorting pathway and with lipid rafts, a type of M domain receptor, which has been suggested to be the sites for effective concentration of Gag. The review provides a summary of these data and discusses the likely direction of future studies. PMID- 15043209 TI - Transmission and immunopathogenesis of FIV in cats as a model for HIV. AB - The feline immunodeficiency virus (FIV) model provides a system to study lentivirus transmission, virus kinetics, pathogenesis, host responses, and immune dysfunction in a natural, out-bred host, under controlled conditions with specific-pathogen-free animals. The diversity of primary FIV strains can be exploited to mirror the range of disease manifestations associated with HIV infection. FIV is infectious via intravenous, intraperitoneal, intradermal, or subcutaneous injection as well as by atraumatic instillation onto the oral, vaginal, or rectal mucosa. Together, these features allow investigators to model specific aspects of HIV infection in a highly relevant and relatively inexpensive animal model. Well-developed areas of the FIV model include: (1) transmission of cell-associated as well as cell-free virus; (2) mucosal infectivity and immunopathogenesis; (3) vertical transmission; (4) acquired immunodeficiency including defects of the innate immune system; (5) thymic dysfunction; (6) neurotropism and neuropathogenesis; (7) host-virus interactions and the role of specific gene products; (8) efficacy of antiviral therapy; and (9) efficacy and immune correlates of experimental vaccines. This review will encompass areas specific to transmission and immunopathogenesis. PMID- 15043210 TI - How HIV evades CTL recognition. AB - The human immunodeficiency virus (HIV) is unique in its capacity to produce chronic disease in almost all infected hosts. To accomplish this, it has evolved multiple mechanisms to effectively evade the immune response. HIV encodes at least one protein that makes infected cells resistant to CTL killing by downmodulating epitope (peptide plus MHC-I protein) density on the infected cell surface. In addition, HIV encodes several mechanisms to promote apoptosis of HIV specific CTLs. The end result is that infected cells have a reduced susceptibility to CTLs, survive longer and produce more viral antigenic variants that can further evade the immune response. PMID- 15043211 TI - In vivo analysis of Nef function. AB - The nef gene is present in all primate lentiviruses (HIV-1, HIV-2, and SIVs). In vivo, Nef has been shown to be a major determinant of virus pathogenicity. In vitro, many different Nef activities have been reported, including CD4 and MHC I downregulation, enhanced virion infectivity, and T-cell activation. These four different activities have been extensively investigated and appear to increase the pathogenicity of the virus. However, the contribution that these activities (individually or together) make to the in vivo phenotype has not been elucidated. The mechanism(s) by which Nef modulates distinct host cell properties has provided great insights into the intricate interaction between virus and host. In this manuscript, we review the different model systems that have been used to study Nef function in vivo and the information that they have provided regarding the best characterized in vitro Nef activities. The knowledge that has been accumulated has provided clues to our understanding of Nef function but they have also left us with many unanswered questions that should be the focus of future in vivo analysis of Nef function. PMID- 15043212 TI - Update on D-ala-peptide T-amide (DAPTA): a viral entry inhibitor that blocks CCR5 chemokine receptors. AB - Peptide T, named for its high threonine content (ASTTTNYT), was derived by a database search which assumed that a relevant receptor binding epitope within env (gp120) would have sequence homology to a known signaling peptide. Binding of radiolabeled gp120 to brain membranes was displaced by peptide T and three octapeptide analogs (including "DAPTA", Dala1-peptide T-amide, the protease resistant analog now in Phase II clinical trials) with the same potency that these four octapeptides blocked infectivity of an early passage patient isolate. This 1986 report was controversial due to a number of laboratories' failure to find peptide T antiviral effects; we now know that peptide T is a potent HIV entry inhibitor selectively targeting CCR5 receptors with minimal effects on the X4 tropic lab adapted virus exclusively in use at that time. Early clinical trials, which demonstrated lack of toxicity and focused on neurological and neurocognitive benefits, are reviewed and data from a small ongoing Phase II trial--the first to assess peptide T's antiviral effects--are presented. Studies using infectivity, receptor binding, chemotaxis, and blockade of gp120-induced neurotoxicity in vitro and in vivo are reviewed, discussed and presented here. Peptide T and analogs of its core pentapeptide, present near the V2 stem of numerous gp120 isolates, are potent ligands for CCR5. Clinical data showing peptide T's immunomodulation of plasma cytokine levels and increases in the percentage of IFNgamma secreting CD8+ T cells in patients with HIV disease are presented and suggests additional therapeutic mechanisms via regulation of specific immunity. PMID- 15043213 TI - Measuring the infectiousness of persons with HIV-1: opportunities for preventing sexual HIV-1 transmission. AB - Methods to reduce sexual transmission of HIV-1 are urgently needed to slow the global HIV-1 epidemic. These methods should include interventions that minimize susceptibility in uninfected populations at risk, as well as interventions that decrease the infectiousness of HIV-1 infected individuals. Surprisingly few interventions to prevent HIV-1 transmission have been targeted at persons who are already infected, although such interventions could have a significant impact on population-wide HIV-1 spread. This review summarizes current knowledge of factors that influence HIV-1 infectiousness, with special attention on the role of genital tract HIV-1 as a surrogate marker of infectiousness. Epidemiologic studies have demonstrated good agreement between factors associated with increased HIV-1 transmission and increased HIV-1 genital tract shedding. Treatment of sexually transmitted diseases and initiation of antiretroviral therapy have been shown to reduce HIV-1 shedding and thus are promising interventions that may reduce HIV-1 transmission. Unresolved issues related to optimal measurement techniques for genital HIV-1, promises and limitations of antiretroviral therapy and vaccines to reduce infectiousness, and other intervention strategies still in development are reviewed. PMID- 15043215 TI - HIV vaccine development: lessons from the past and promise for the future. AB - The global HIV epidemic continues to expand, exceeding previous predictions and causing tremendous suffering. An effective vaccine represents the best hope to curtail the HIV epidemic. The past fifteen years of HIV vaccine clinical trials have not identified an ideal HIV vaccine, but have provided many valuable lessons that contribute to the current generation of promising HIV vaccine regimens. An enhanced understanding of HIV and SIV immunopathogenesis has facilitated the design of vaccination regimens that elicit specific immune responses and effector mechanisms. Intensive investigation of recombinant gp120 subunit vaccines has revealed a previously unexpected complexity in eliciting neutralizing antibodies that are active against primary isolate viruses. The importance of CD8+ CTL responses in controlling HIV and SIV viremia has led to a series of vaccine candidates that effectively induce these responses. Proof that vaccination can prevent SIV/HIV disease has now been obtained in simian models of AIDS. A number of promising HIV vaccine regimens are currently being evaluated in human trials, and the pipeline of new vaccine vectors and combination regimens appears robust. Although challenges to the development of a safe and effective global HIV vaccine remain, the outlook for HIV vaccines in the future is bright. PMID- 15043214 TI - Sugar and spice: viral envelope-DC-SIGN interactions in HIV pathogenesis. AB - DC-SIGN is a calcium dependent lectin that binds to HIV envelope, gp120, with high affinity. Its expression on dendritic cells, coupled with its ability to facilitate the binding and subsequent transfer of virions to permissive T-cells, has led to the hypothesis that DC-SIGN may serve as a conduit the transfer of HIV from the peripheral mucosa to secondary lymphoid organs. Studies have shown that DC-SIGN bound virions can maintain their infectivity for prolonged periods of time despite evidence that DC-SIGN itself may serve as an antigen receptor. How HIV subverts the normal function of DC-SIGN to establish a primary infection in the host is unclear. Therefore, understanding the structural and immunological basis for DC-SIGN's function will help us realize the role that DC-SIGN may play in viral transmission and pathogenesis. Importantly, DC-SIGN/envelope interactions may represent a new target for microbicide and vaccine development efforts. Here, we review recent studies on DC-SIGN's structure and function in an effort to present testable models of DC-SIGN's role in HIV pathogenesis. PMID- 15043216 TI - Rodent models for HIV-1 infection and disease. AB - The development of a predictive, small animal model for human immunodeficiency virus type 1 (HIV-1) disease would greatly facilitate the analysis of many aspects of viral infection, pathogenesis and treatment. While numerous small animal models exist which emulate various aspects of HIV-1 infection and/or disease in humans, none of these models support robust HIV-1 replication within the context of an intact immune system. Despite this major limitation, these models have helped to elucidate different aspects of HIV-1 pathogenesis in humans. Moreover, recent advances regarding the underlying nature of the blocks to viral replication in non-human cells have raised the possibility that rodents may be engineered to support HIV-1 infection. This review will focus on recent attempts to develop a rodent model for HIV-1 disease, and will also describe currently available systems for studying HIV-1. PMID- 15043217 TI - WebQTL: web-based complex trait analysis. AB - WebQTL is a website that combines databases of complex traits with fast software for mapping quantitative trait loci (QTLs) and for searching for correlations among traits. WebQTL also includes well-curated genotype data for five sets of mouse recombinant inbred (RI) lines. Thus, to identify QTLs, users need provide only quantitative trait data from one of the supported populations. The WebQTL databases include both biological traits--neuroanatomical, pharmacological, and behavioral traits--and microarray-based gene expression data from BXD RI lines. A search function finds correlations between RNA expression and biological traits, and mapping functions find QTLs for either type of trait. The WebQTL service is available at http://www.webqtl.org/. PMID- 15043218 TI - EMAP and EMAGE: a framework for understanding spatially organized data. AB - The Edinburgh MouseAtlas Project (EMAP) is a time-series of mouse-embryo volumetric models. The models provide a context-free spatial framework onto which structural interpretations and experimental data can be mapped. This enables collation, comparison, and query of complex spatial patterns with respect to each other and with respect to known or hypothesized structure. The atlas also includes a time-dependent anatomical ontology and mapping between the ontology and the spatial models in the form of delineated anatomical regions or tissues. The models provide a natural, graphical context for browsing and visualizing complex data. The Edinburgh Mouse Atlas Gene-Expression Database (EMAGE) is one of the first applications of the EMAP framework and provides a spatially mapped gene-expression database with associated tools for data mapping, submission, and query. In this article, we describe the underlying principles of the Atlas and the gene-expression database, and provide a practical introduction to the use of the EMAP and EMAGE tools, including use of new techniques for whole body gene expression data capture and mapping. PMID- 15043219 TI - Informatics center for mouse genomics: the dissection of complex traits of the nervous system. AB - In recent years, there has been an explosion in the number of tools and techniques available to researchers interested in exploring the genetic basis of all aspects of central nervous system (CNS) development and function. Here, we exploit a powerful new reductionist approach to explore the genetic basis of the very significant structural and molecular differences between the brains of different strains of mice, called either complex trait or quantitative trait loci (QTL) analysis. Our specific focus has been to provide universal access over the web to tools for the genetic dissection of complex traits of the CNS--tools that allow researchers to map genes that modulate phenotypes at a variety of levels ranging from the molecular all the way to the anatomy of the entire brain. Our website, The Mouse Brain Library (MBL; http://mbl.org) is comprised of four interrelated components that are designed to support this goal: The Brain Library, iScope, Neurocartographer, and WebQTL. The centerpiece of the MBL is an image database of histologically prepared museum-quality slides representing nearly 2000 mice from over 120 strains--a library suitable for stereologic analysis of regional volume. The iScope provides fast access to the entire slide collection using streaming video technology, enabling neuroscientists to acquire high-magnification images of any CNS region for any of the mice in the MBL. Neurocartographer provides automatic segmentation of images from the MBL by warping precisely delineated boundaries from a 3D atlas of the mouse brain. Finally, WebQTL provides statistical and graphical analysis of linkage between phenotypes and genotypes. PMID- 15043220 TI - Genetic correlates of gene expression in recombinant inbred strains: a relational model system to explore neurobehavioral phenotypes. AB - Full genome sequencing, high-density genotyping, expanding sets of microarray assays, and systematic phenotyping of neuroanatomical and behavioral traits are producing a wealth of data on the mouse central nervous system (CNS). These disparate resources are still poorly integrated. One solution is to acquire these data using a common reference population of isogenic lines of mice, providing a point of integration between the data types. Recombinant inbred (RI) mice, derived through inbreeding of progeny from an inbred cross, are a powerful tool for complex trait mapping and analysis of the challenging phenotypes of neuroscientific interest. These isogenic RI lines are a retrievable genetic resource that can be repeatedly studied using a wide variety of assays. Diverse data sets can be related through fixed and known genomes, using tools such as the interactive web-based system for complex trait analysis, www.WebQTL.org. In this report, we demonstrate the use of WebQTL to explore complex interactions among a wide variety of traits--from mRNA transcripts to the impressive behavioral and pharmacological variation among RI strains. The relational approach exploiting a common set of strains facilitates study of multiple effects of single genes (pleiotropy) without a priori hypotheses required. Here we demonstrate the power of this technique through genetic correlation of gene expression with a database of neurobehavioral phenotypes collected in these strains of mice through more than 20 years of experimentation. By repeatedly studying the same panel of mice, early data can be re-examined in light of technological advances unforeseen at the time of their initial collection. PMID- 15043221 TI - A guide to building image-centric databases. AB - There is a paucity of image-centric neuroinformatics infrastructure within the individual investigator's laboratory despite the obvious need for automation and integration of experimental results. Yet, solutions can often be readily built using off-the-shelf databases and associated tools. Doing so simplifies day-to day research operation and increases throughput. Proper construction of in-house solutions may also expedite community-wide integration of private and public data repositories. Here we describe neuroinformatics approaches at different levels of functionality, required expertise, and size of image datasets. The simplest approach offers ease of image browsing and rudimentary searching. More sophisticated systems provide powerful search capabilities, a means of tracking analysis, and even automated serial processing pipelines. In this practicum, we provide guidance in selecting among the different options. PMID- 15043222 TI - The cell-centered database: a database for multiscale structural and protein localization data from light and electron microscopy. AB - The creation of structured shared data repositories for molecular data in the form of web-accessible databases like GenBank has been a driving force behind the genomic revolution. These resources serve not only to organize and manage molecular data being created by researchers around the globe, but also provide the starting point for data mining operations to uncover interesting information present in the large amount of sequence and structural data. To realize the full impact of the genomic and proteomic efforts of the last decade, similar resources are needed for structural and biochemical complexity in biological systems beyond the molecular level, where proteins and macromolecular complexes are situated within their cellular and tissue environments. In this review, we discuss our efforts in the development of neuroinformatics resources for managing and mining cell level imaging data derived from light and electron microscopy. We describe the main features of our web-accessible database, the Cell Centered Database (CCDB; http://ncmir.ucsd.edu/CCDB/), designed for structural and protein localization information at scales ranging from large expanses of tissue to cellular microdomains with their associated macromolecular constituents. The CCDB was created to make 3D microscopic imaging data available to the scientific community and to serve as a resource for investigating structural and macromolecular complexity of cells and tissues, particularly in the rodent nervous system. PMID- 15043223 TI - The informatics of a C57BL/6J mouse brain atlas. AB - The Mouse Atlas Project (MAP) aims to produce a framework for organizing and analyzing the large volumes of neuroscientific data produced by the proliferation of genetically modified animals. Atlases provide an invaluable aid in understanding the impact of genetic manipulations by providing a standard for comparison. We use a digital atlas as the hub of an informatics network, correlating imaging data, such as structural imaging and histology, with text based data, such as nomenclature, connections, and references. We generated brain volumes using magnetic resonance microscopy (MRM), classical histology, and immunohistochemistry, and registered them into a common and defined coordinate system. Specially designed viewers were developed in order to visualize multiple datasets simultaneously and to coordinate between textual and image data. Researchers can navigate through the brain interchangeably, in either a text based or image-based representation that automatically updates information as they move. The atlas also allows the independent entry of other types of data, the facile retrieval of information, and the straight-forward display of images. In conjunction with centralized servers, image and text data can be kept current and can decrease the burden on individual researchers' computers. A comprehensive framework that encompasses many forms of information in the context of anatomic imaging holds tremendous promise for producing new insights. The atlas and associated tools can be found at http://www.loni.ucla.edu/MAP. PMID- 15043224 TI - Sharing the mighty mouse. PMID- 15043225 TI - The incredible career of Peter J. Safar, MD: the Michelangelo of acute medicine. PMID- 15043226 TI - Pioneering contributions of Peter Safar to intensive care and the founding of the Society of Critical Care Medicine. PMID- 15043227 TI - Controlled normothermia in neurologic intensive care. AB - Preclinical studies of cerebral ischemia and trauma find increased brain tissue injury and worsened functional outcomes if the brain temperature exceeds 39 degrees C. Several retrospective studies of patients with new-onset stroke, intracerebral hemorrhage, or subarachnoid hemorrhage support these observations. However, fever is very common among these patients early after the onset of their disease, particularly if they are in the ICU for a week or more, and brain temperatures are likely to be as much as 2 degrees C higher than rectal temperatures. Finally, intravascular temperature modulation has been shown to be more effective for preventing fever than conventional methods, such as antipyretic medications or surface-cooling techniques. Further study is needed to establish if such better control of temperature will lead to improved outcomes. PMID- 15043228 TI - Suspended animation for resuscitation from exsanguinating hemorrhage. AB - In dogs, isotonic saline at 0-4 degrees C, flushed into the aorta at a rate of 1 2 L/min, with drainage of the vena cava, can achieve deep to profound hypothermia of vital organs at a cooling rate of up to 3 degrees C per minute. This achieves preservation of viability of the organism during predictable durations of no flow: cardiac arrest of 15-20 mins at Tty of 30-35 degrees C, cardiac arrest of 30 mins at Tty of 25 degrees C, cardiac arrest of 60 mins at Tty of 15 degrees C, and cardiac arrest of 90 mins at Tty of 10 degrees C. So far, pharmacologic approaches have not resulted in any breakthrough effect on outcome above that achieved with hypothermia, except perhaps the antioxidant tempol. Additional studies of novel drugs and, perhaps, combination therapies remain warranted. The optimal fluids to have in the circulation during circulatory arrest and reperfusions need to be determined. As laboratory studies to optimize suspended animation proceed, clinical trials should be initiated. In addition, devices should be developed to facilitate induction of suspended animation, eventually in the field. PMID- 15043229 TI - Smart aortic arch catheter: moving suspended animation from the laboratory to the field. PMID- 15043230 TI - From Resusci-Anne to Sim-Man: the evolution of simulators in medicine. PMID- 15043231 TI - National Medical Simulation training program in Denmark. PMID- 15043232 TI - Improving medical crisis team performance. PMID- 15043233 TI - Experience with medical student simulation education. PMID- 15043234 TI - Simulation in medical students' critical thinking. PMID- 15043235 TI - Pediatric simulation: a valuable tool for pediatric medical education. PMID- 15043236 TI - The law of unintended consequences. PMID- 15043237 TI - "Physicians' estimates of cardiac index and intravascular volume based on clinical assessment versus transesophageal Doppler measurements obtained by critical care nurses". PMID- 15043238 TI - Nocturnal care interactions with patients in critical care units. AB - BACKGROUND: Sleep deprivation is common in critically ill patients and may have long-term effects on health outcomes and patients' morbidity. Clustering nocturnal care has been recommended to improve patients' sleep. OBJECTIVES: To (1) examine the frequency, pattern, and types of nocturnal care interactions with patients in 4 critical care units; (2) analyze the relationships among these interactions and patients' variables (age, sex, acuity) and site of admission to the intensive care unit; and (3) analyze the differences in patterns of nocturnal care activities among the 4 units. METHODS: A randomized retrospective review of the medical records of 50 patients was used to record care activities from 7 PM to 7 AM in 4 critical care units. RESULTS: Data consisted of interactions during 147 nights. The mean number of care interactions per night was 42.6 (SD 11.3). Interactions were most frequent at midnight and least frequent at 3 AM. Only 9 uninterrupted periods of 2 to 3 hours were available for sleep (6% of 147 nights studied). Frequency of interactions correlated significantly with patients' acuity scores (r = 0.32, all Ps < .05). A sleep-promoting intervention was documented for only 1 of the 147 nights, and 62% of routine daily baths were provided between 9 PM and 6 AM. CONCLUSIONS: The high frequency of nocturnal care interactions left patients few uninterrupted periods for sleep. Interventions to expand the period around 3 AM when interactions are least common could increase opportunities for sleep. PMID- 15043239 TI - Pain levels experienced with activities after cardiac surgery. AB - BACKGROUND: Acute pain is common after cardiac surgery and can keep patients from participating in activities that prevent postoperative complications. Accurate assessment and understanding of pain are vital for providing satisfactory pain control and optimizing recovery. OBJECTIVES: To describe pain levels for 5 activities expected of patients after cardiac surgery on postoperative days 1 to 6 and changes in pain levels after chest tube removal and extubation. METHODS: Adults who underwent cardiac surgery were asked to rate the pain associated with various types of activities on postoperative days 1 to 6. Pain levels were compared by postoperative day, activity, and type of cardiac surgery. Pain scores before and after chest tube removal and extubation also were analyzed. RESULTS: Pain scores were higher on earlier postoperative days. The order of overall pain scores among activities (P < .01) from highest to lowest was coughing, moving or turning in bed, getting up, deep breathing or using the incentive spirometer, and resting. Changes in pain reported with coughing (P = .03) and deep breathing or using the incentive spirometer (P = .005) differed significantly over time between surgery groups. After chest tubes were discontinued, patients had lower pain levels at rest (P = .01), with coughing (P = .05), and when getting up (P = .03). CONCLUSIONS: Pain relief is an important outcome of care. A comprehensive, individualized assessment of pain that incorporates activity levels is necessary to promote satisfactory management of pain. PMID- 15043240 TI - Pain assessment and management in critically ill intubated patients: a retrospective study. AB - BACKGROUND: Little research has been done on pain assessment in critical care, especially in patients who cannot communicate verbally. OBJECTIVES: To describe (1) pain indicators used by nurses and physicians for pain assessment, (2) pain management (pharmacological and nonpharmacological interventions) undertaken by nurses to relieve pain, and (3) pain indicators used for pain reassessment by nurses to verify the effectiveness of pain management in patients who are intubated. METHODS: Medical files from 2 specialized healthcare centers in Quebec City, Quebec, were reviewed. A data collection instrument based on Melzack's theory was developed from existing tools. Pain-related indicators were clustered into nonobservable/subjective (patients' self-reports of pain) and observable/objective (physiological and behavioral) categories. RESULTS: A total of 183 pain episodes in 52 patients who received mechanical ventilation were analyzed. Observable indicators were recorded 97% of the time. Patients' self reports of pain were recorded only 29% of the time, a practice contradictory to recommendations for pain assessment. Pharmacological interventions were used more often (89% of the time) than nonpharmacological interventions (<25%) for managing pain. Almost 40% of the time, pain was not reassessed after an intervention. For reassessments, observable indicators were recorded 66% of the time; patients self reports were recorded only 8% of the time. CONCLUSIONS: Pain documentation in medical files is incomplete or inadequate. The lack of a pain assessment tool may contribute to this situation. Research is still needed in the development of tools to enhance pain assessment in critically ill intubated patients. PMID- 15043241 TI - Illness representation after acute myocardial infarction: impact on in-hospital recovery. AB - BACKGROUND: Despite significant progress in the treatment of coronary artery disease, myocardial infarction is still the leading cause of death in the United States. As suggested by Leventhal's Self-Regulation Model of Illness, the continued high morbidity and mortality may be due to a failure to address the role of psychosocial factors such as illness representation, depression, and anxiety in recovery. OBJECTIVE: To determine the relationship between illness representation of myocardial infarction and the occurrence of in-hospital complications and if anxiety and depression mediate this relationship. METHOD: A prospective correlational design was used to measure illness representation, depression, and anxiety 24 to 48 hours after admission for myocardial infarction in 49 patients and the frequency of complications during the acute event. Logistic regression was used to determine the likelihood of experiencing a complication. RESULTS: When demographic and clinical variables were controlled for, the more negative the representation of illness, the greater were the odds of experiencing a complication (chi2 = 16.9, df = 6, P =.01). The odds of experiencing a complication increased 5.1% for each 1 unit increase in the score on the Illness Preparation Questionnaire (B = 0.05, Wald = 4.442, Exp(B) = 1.051, 95% CI = 1.003-1.1010). Neither anxiety (chi2 = 3.0, df = 1, P =. 09) nor depression (chi2 = 2.5, df = 1, P = .11) were significant predictors of the occurrence of complications. CONCLUSION: In these patients, illness representation was predictive of the likelihood of experiencing a complication. Thus, illness representation appears to be an important psychosocial factor in acute recovery from myocardial infarction. PMID- 15043242 TI - Assessment of anxiety in intensive care patients by using the Faces Anxiety Scale. AB - BACKGROUND: Anxiety is difficult to detect in patients receiving mechanical ventilation because clinical signs are confounded and patients often cannot respond to validated anxiety measures. Most patients can respond to the single item Faces Anxiety Scale. OBJECTIVES: To assess the validity of the Faces Anxiety Scale, the frequency and severity of state anxiety, and correlates of anxiety in intensive care patients. METHODS: A research assistant made a single clinical judgment of anxiety in the range of 1 to 10 on the basis of patients' nonverbal responses (ie, nods) to 9 questions about mood and their physical and behavioral signs. Patients then responded to the Faces Anxiety Scale. Demographic, clinical, and pharmacological data were obtained from the patients' charts. RESULTS: Mean age of the 106 patients was 61 years; 62% were men. Admission diagnoses were cardiovascular in 26% of patients, respiratory in 26%, trauma in 18%, neurological in 12%, gastrointestinal in 12%, and other in 6%. At the time of anxiety assessment, 89% were receiving mechanical ventilation. The correlation between patients' self-reports of anxiety on the Anxiety Faces Scale and the research assistant's assessments was 0.64 (P <.001). Some anxiety was reported by 85% of patients (mean level 2.9; SD 1.2). Anxiety levels were lower in patients who had recently received sedatives or opioids but were not related to heart rate or blood pressure. CONCLUSIONS: The Faces Anxiety Scale is a valid means of measuring anxiety in intensive care patients. Anxiety is common in these patients and is often moderate to severe. PMID- 15043243 TI - Relationship of age and sex to health-related quality of life in patients with heart failure. AB - BACKGROUND: Although health-related quality of life is diminished among patients with chronic heart failure, few investigators have examined interactions of age and sex with health-related quality of life longitudinally. OBJECTIVES: To examine differences in health-related quality of life among 4 groups of patients with heart failure on the basis of age (< 65 years and > or = 65 years) and sex and to evaluate relationships of age and sex to changes in health-related quality of life during 6 months. METHODS: Patients from 2 outpatient clinics in an urban county hospital were interviewed at baseline and 26 weeks later Health-related quality of life was measured by using the Minnesota Living With Heart Failure Questionnaire and the Chronic Heart Failure Questionnaire. RESULTS: A total of 165 patients (52% women; mean age, 57.6 years) completed interviews at baseline and 26 weeks later. At baseline, patients younger than 65 years had poorer health related quality of life scores on total scales and some subscales than did older patients. Women had poorer scores than did men on some scales, particularly the emotional subscales. At 26 weeks, patients younger than 65 had poorer total health-related quality of life on 1 scale than did patients 65 and older, and women had poorer scores than did men on 1 total scale. With demographic and clinical factors controlled for, women younger than 65 had improvements in health related quality of life on some scales. CONCLUSIONS: Women younger than 65 years had relatively poorer initial health-related quality of life that improved after 26 weeks. PMID- 15043244 TI - The resurrection of spironolactone on its golden anniversary. AB - The edematous states, specifically those in CHF and cirrhosis of the liver, are associated with excessive aldosterone secretion and represent states of secondary hyperaldosteremia. Aldosterone promotes sodium retention by the renal tubules. Spironolactone, first introduced 50 years ago (1953), blocks the action of aldosterone on renal transport of electrolytes, thus acting as an effective diuretic, and in addition, has potentiating effects on other diuretics, including the thiazides. Spironolactone has undesirable side effects that have limited its clinical use; the most significant are impotence, gynecomastia, and hirsutism. Eplerenone, a recently introduced selective ARA, decreases morbidity and mortality in patients with CHF following MI and has none of the androgenic or estrogenic side effects of spironolactone. Eplerenone is an effective alternative for spironolactone. PMID- 15043245 TI - Nonischemic ST-segment elevation. PMID- 15043246 TI - "Evaluation of chemical dot thermometers in orally intubated patients". PMID- 15043247 TI - Thermodynamics and kinetics of solute transfer in reversed-phase liquid chromatography. Effect of annelation in polycyclic aromatic hydrocarbons. AB - A series of four-ring polycyclic aromatic hydrocarbons (PAHs) with varying annelation structure was studied by reversed-phase liquid chromatography. Using a polymeric octadecylsilica stationary phase over a temperature range from 273 to 303 K and an average pressure range from 585 to 3585 psi (1 psi = 6894.76 Pa), the thermodynamic and kinetic aspects of the retention mechanism were examined. Thermodynamic behavior was characterized by the retention factor, together with the associated changes in molar enthalpy and molar volume, whereas kinetic behavior was characterized by the rate constants, together with the associated activation enthalpies and activation volumes. The data indicate that pyrene, with a more condensed annelation structure, exhibits smaller changes in molar enthalpy and molar volume (delta Hsm = -4.4 kcal/mol, delta Vsm = -1.9 ml/mol; 1 cal = 4.184J) than PAHs with a more linear structure such as chrysene (delta Hsm = -8.2 kcal/mol, delta Vsm = - 11.7 ml/mol). The kinetic data indicate that pyrene undergoes faster rates of transport than chrysene (k(ms) = 313 and 14 s(-1), respectively), but the non-planar benzo[c]phenanthrene undergoes the fastest transport (k(ms) = 330 s(-1)). The activation enthalpies and activation volumes are similarly affected by the annelation structure. It is noteworthy that deviations from the exponentially modified Gaussian (EMG) model are observed for some PAH zone profiles at the lowest temperature, which suggests a possible change in retention mechanism. In order to characterize these deviations, the non linear chromatography (NLC) model and a new bi-exponentially modified Gaussian (E2MG) model were examined. The regression results indicate that neither the NLC nor E2MG model offer significant improvements in the statistical quality of fit or provide a better description of the observed retention behavior. PMID- 15043248 TI - Analysis of the mechanism of retention on graphitic carbon by a computational chemical method. AB - Retention mechanism on a graphitic carbon was analyzed by computational chemical calculation. The model graphitic carbon phase was a large polycyclic aromatic hydrocarbon (PAH) and analytes were carbohydrates and hydrocarbons separated by liquid and gas chromatography. Molecular mechanics calculation was fast and suggested their retention order and main retention force. Molecular orbital package calculation (MOPAC) demonstrated their complex form. PMID- 15043249 TI - Elucidation of retention mechanisms on hypercrosslinked polystyrene used as column packing material for high-performance liquid chromatography. AB - Establishing of basic retention mechanisms was considered the key target during the development of new column packing materials. To extract, from an appropriate retention data matrix on hypercrosslinked polystyrene Chromalite 5HGN, certain factors that can be brought in an obvious correspondence with known retention mechanisms, the principal component analysis (PCA) was applied. The approach was used to elucidate the adsorption properties of the above novel HPLC packing. Besides HPLC, knowledge of retention mechanisms helps to reveal perspective application area for the hypercrosslinked polystyrene-type materials in solid phase extraction (SPE) and low-pressure preparative LC. PMID- 15043250 TI - Comparison of equations describing band broadening in high-performance liquid chromatography. AB - Several models are well established that describe band broadening in gas and liquid chromatography, including those due to Van Deemter and Knox. Comparison of competing models is complicated if raw data are noisy or if the equations to be fitted to data contain many adjustable parameters. This paper describes a comparison of fitting the Van Deemter, Knox and other equations to low noise data gathered during the separation of propyl- and methylparaben by HPLC. Equations are compared using established statistical methods, including analysis of residuals, inference of parameter estimates and Akaikes Information Criterion for model identification. This work indicates that equations that account for non linear band broadening at elevated mobile phase velocities are more successful at describing the relationship between height equivalent to a theoretical plate, H, and the velocity of the mobile phase, u. PMID- 15043251 TI - Stationary-phase effects in gradient high-performance liquid chromatography. AB - The type of the stationary phase for reversed-phase liquid chromatography significantly affects the sample polarity range that can be covered using gradients of organic solvents in water. The polarity range available for gradient separations of samples containing compounds differing in the lipophilic parts of the molecules can be characterized by "gradient lipophilic capacity", Pl, based on the retention of standard compounds with a repeat lipophilic structural unit, such as a methylene group. The gradient lipophilic capacity is also suitable to characterize the separation possibilities of the columns in non-aqueous reversed phase gradient elution of strongly non-polar compounds, such as triacylglycerols. In the same way, the suitability of various columns for reversed-phase gradient separations of oligomers can be characterized by "gradient oligomer capacity", as demonstrated in the example of oligo(ethylene glycols). To enable a comparison of the properties of stationary phases independent of column efficiency and dimensions, the gradient lipophilic capacity or the gradient oligomer capacity should be normalized for a "standard" column plate number, gradient range and volume (in column hold-up volume units). The gradient lipophilic capacity or the gradient oligomer capacity and the number of compounds that can be resolved during a gradient run decrease as the initial concentration of the strong solvent in the mobile phase increases and (or) the gradient time decreases. These quantities can be used to select a suitable column and to adjust the optimum gradient profile (the initial composition of the mobile phase and the gradient steepness) with respect to the time of analysis and the number of oligomers or other compounds with regular repeat structural groups that can be resolved during the gradient run. PMID- 15043252 TI - Optimization strategies for the analysis and purification of drug discovery compounds by reversed-phase high-performance liquid chromatography with high-pH mobile phases. AB - Careful selection of both high-pH mobile phase as well as organic modifier, was performed in order to develop and optimize HPLC conditions for the separation of drug discovery compounds. High-pH mobile phases provide excellent chromatographic resolution and increased mass loading of basic compounds. The analytical methods so defined have been successfully transferred to preparative automated UV directed purification, an important fact due to the increasing number of samples requiring purification. It should be noted that, the single prerequisite for this approach is an analytical LC-UV-MS run, therefore the system has the ability to collect only fractions likely to contain the target product. A cost-effective strategy for maximizing the purification of drug discovery compounds is proposed. PMID- 15043253 TI - Computational study of the band broadening in two-dimensional etched packed bed columns for on-chip high-performance liquid chromatography. AB - The chromatographic performance of several straightforward two-dimensional etched packed bed column lay-outs (equilaterally staggered arrays of, respectively, circular, hexagonal, and diamond-like pillars) has been compared using commercial computational fluid dynamics software. In all cases, the bed porosity was kept at epsilon = 0.4 and a retained component with zone capacity ratio k" = 2 was considered. Exploring the use of six different possible characteristic dimensions to bring the Van Deemter plots of the three different considered particle shapes into agreement, none of them yielded a perfect agreement. Using the pillar volume based equivalent cylinder diameter (deq) as the characteristic dimension, the diamond-like pillars yielded a significantly smaller h(min) value than the cylinders and the hexagons (h(min) approximately equal to 0.74 for the former versus h(min) approximately equal to 0.83 for the two latter). Including the flow resistance into the analysis, it was found that the "hydrodynamic" shape of the particles has an important influence on the separation impedance E. The more axially elongated diamond pillars yielded an Emin number as small Emin = 180 (for a retained component with k" = 2), i.e. about 40% smaller than the cylinders and the hexagons (Emin = 300-330). The obtained h(min) and Emin values are also significantly smaller than the values often cited for the best possible packed bed HPLC columns. We believe this is a consequence of the assumed perfect homogeneity of the etched structures, and hence hints at the potential benefits of perfectly ordered chromatographic columns, as was already inferred by Knox [J. Chromatogr. A 831 (1999) 3; 960 (2002) 7] and He et al. [Anal. Chem. 70 (1998) 3790]. PMID- 15043254 TI - Solid-phase extraction of polar compounds with a hydrophilic copolymeric sorbent. AB - A new synthesized copolymer based on N-vinylimidazole-divinylbenzene (VIm-DVB) was tested as a sorbent for the solid-phase extraction (SPE) of polar analytes. In the on-line SPE, this synthesized sorbent enabled 100 ml of sample to be preconcentrated with recoveries as high as 80% for oxamyl, phenol (Ph) and derivates, bentazone and (4-chloro-2-methylphenoxy)acetic acid (MCPA). For the off-line SPE, 1000 ml of sample was extracted and recoveries were higher than 92% for all compounds with the exception of oxamyl (83%) and methomyl (78%). The VIm DVB sorbent gives better recoveries than the previously synthesized 4 vinylpyridine-divinylbenzene (VP-DVB) resin and similar to such highly crosslinked commercial sorbents as LiChrolut EN or Oasis HLB. Real water samples were used to validate the on-line SPE method. Linearity was good and detection limits were between 0.1 and 0.2 microg l(-1). PMID- 15043255 TI - Automated sample preparation based on the sequential injection principle. Solid phase extraction on a molecularly imprinted polymer coupled on-line to high performance liquid chromatography. AB - A molecularly imprinted polymer (MIP) prepared using caffeine, as a template, was validated as a selective sorbent for solid-phase extraction (SPE), within an automated on-line sample preparation method. The polymer produced was packed in a polypropylene cartridge, which was incorporated in a flow system prior to the HPLC analytical instrumentation. The principle of sequential injection was utilised for a rapid automated and efficient SPE procedure on the MIP. Samples, buffers, washing and elution solvents were introduced to the extraction cartridge via a peristaltic pump and a multi-position valve, both controlled by appropriate software developed in-house. The method was optimised in terms of flow rates, extraction time and volume. After extraction, the final eluent from the extraction cartridge was directed to the injection loop and was subsequently analysed on HPLC. The overall set-up facilitated unattended operation, operation and improved both mixing fluidics and method development flexibility. This system may be readily built in the laboratory and can be further used as an automated platform for on-line sample preparation. PMID- 15043256 TI - Comparison of solid-phase microextraction and stir bar sorptive extraction for determining six organophosphorus insecticides in honey by liquid chromatography mass spectrometry. AB - Two approaches based on sorptive extraction, solid-phase microextraction (SPME) and stir bar sorptive extraction (SBSE), in combination with liquid chromatography (LC)-atmospheric pressure chemical ionization mass spectrometry (MS) have been assayed for analyzing chlorpyriphos methyl, diazinon, fonofos, phenthoate, phosalone, and pirimiphos ethyl in honey. In both, SPME and SBSE, enrichment was performed using a poly(dimethylsiloxane) coating. Significant parameters affecting sorption process such as sample volume, sorption and desorption times, ionic strength, elution solvent, and dilution (water/honey) proportion were optimized and discussed. Performance of both methods has been compared through the determination of linearity, extraction efficiencies, and limits of quantification. Relative standard deviations for the studied compounds were from 3 to 10% by SPME and from 5 to 9% by SBSE. Both methods were linear in a range of at least two orders of magnitude, and the limits of quantification reached ranging from 0.04 to 0.4 mg kg(-1) by SBSE, and from 0.8 to 2 mg kg(-1) by SPME. The two procedures were applied for analyzing 15 commercial honeys of different botanical origin. SPME and SBSE in combination with LC-MS enabled a rapid and simple determination of organophosphorus pesticides in honey. SBSE showed higher concentration capability (large quantities of sample can be handled) and greater accuracy (between 5 and 20 times) and sensitivity (between 10 and 50 times) than SPME: thus, under equal conditions, SBSE is the recommended technique for pesticide analysis in honey. PMID- 15043257 TI - Solid-phase microextraction coupled with high-performance liquid chromatography for the analysis of heterocyclic aromatic amines. AB - Solid-phase microextraction (SPME) coupled with high-performance liquid chromatography (HPLC) with UV diode array detection (DAD) for the analysis of heterocyclic aromatic amines (HAs) is described. Four kinds of fiber coatings: Carbowax-templated resin (CW-TPR), Carbowax-divinylbenzene (CW-DVB), poly(dimethylsiloxane)-divinylbenzene (PDMS-DVB) and polyacrylate (PA) were evaluated for extraction of nine most biologically active heterocyclic aromatic amines. Different parameters affecting to the microextraction and determination of HAs were studied, such as absorption and desorption time, desorption mode, composition of the solvent for desorption, pH, ionic strength, and percentage of methanol in the sample. To determine these amines in food samples a new simplified procedure is proposed, consisting of treatment of the sample with methanolic NaOH prior microextraction by CW-TPR fiber coating and HPLC-DAD determination. The advantages of this new method are the reduced amounts of time and organic solvents required. PMID- 15043258 TI - Evaluation of a novel anion-exchange restricted-access sorbent for on-line sample clean-up prior to the determination of acidic compounds in plasma by liquid chromatography. AB - A new kind of silica-based restricted-access material (RAM) with anionic properties has been tested in pre-columns for on-line solid-phase extraction of acidic compounds from directly injected plasma samples prior to their determination by reversed-phase liquid chromatography (LC), using the column switching technique. The outer surface of the porous RAM particles contains hydrophilic diol groups while diethylaminoethyl (DEAE) groups are bound to the internal surface which gives the sorbent the properties of a weak anion exchanger towards low-molecular-mass compounds. Due to an appropriate pore diameter (about 6 nm), macromolecules, such as proteins, are physically excluded from the pores and flushed directly out during the sample clean-up process, while small compounds have access to the inner surface and can be retained mainly by electrostatic interactions. The retention capability of this novel packing material has been tested for some hydrophilic acidic compounds such as aspartic acid, glutamic acid, ascorbic acid and acetylcysteine as well as for some more hydrophobic drugs such as naproxen, ibuprofen and diclofenac, used as model compounds. The influence of the composition of the washing liquid on the retention of the analytes in the pre-column has been investigated. The efficiency of the sorbent to clean-up complex matrices was also tested using human plasma and urine samples. A generic washing liquid composition was then selected in order to obtain efficient and selective sample clean-up as well as a high recovery of the acidic analytes. PMID- 15043259 TI - Development of an analytical method for polycyclic aromatic hydrocarbons and their derivatives. AB - The possibilities of utilising pressurised liquid extraction for five nitro polycyclic aromatic hydrocarbons from an inert matrix are shown. Different extraction temperatures and pressures were tested. The highest recoveries were obtained at extraction pressure 14 MPa and temperature 100 C. Separation of non polar, aromatic and polar fractions by the silica gel column chromatography is shown. n-Hexane, cyclohexane and dichloromethane as a solvent were tested. The best separations of monitored fractions were obtained, when extract was dissolved in cyclohexane. Non-polar and aromatic fractions eluted together when the extract was dissolved in dichloromethane. PMID- 15043260 TI - Potential of coacervation processes for the extraction of amphiphiles (linear alkyl benzenesulphonates) from sewage sludge samples prior to liquid chromatography. AB - A new approach was developed for the monitoring of linear alkyl (C10-C13) benzenesulphonates (LASs) in sewage sludge. It was based on their extraction with the anionic surfactant sodium dodecanesulphonate (SDoS) that undergoes coacervation under acid conditions. The target compounds formed mixed aggregates with SDoS by ideal hydrophobic interactions which made possible the breakdown of LAS-sludge interactions and provided high extraction yields. Variables affecting extraction were optimised using a fortified dehydrated sludge. Recoveries for LASs were found independent on the length of alkyl chain. Liquid chromatography fluorimetry was used for separation and detection of LAS homologues. Detection limit for LAS in the sludge was 5 mg/kg. Concentration levels of total LASs in activated and dehydrated sludge collected from two different sewage treatment plants were in the range 0.26-0.56 g/kg with LAS homologues ranging from 29 to 223 mg/kg. The method did not require clean-up or preconcentration steps. PMID- 15043261 TI - Selective removal of DNA from protein solution with copolymer particles derived from N,N-dimethylaminopropylacrylamide. AB - To remove nucleic acids from cellular products as drugs, cross-linked N,N dimethylaminopropylacrylamide (DMP) particles with cationic functional groups were prepared. The particle's hydrophobicity and its anion-exchange capacity were easily adjusted by changing the cross-linking agent and the DMP ratio in the cross-linking, respectively. When divinylbenzene (DVB) was used as a cross linking agent and the DMP ratio (in the cross-linking) was adjusted to 90 mol%, the particles (DMP-DVB, 90:10) showed the highest adsorbing activity of DNA (salmon spermary). Its adsorption capacity was 54 mg/ml adsorbent. On the other hand, the adsorption of bovine serum albumin (BSA) to the DMP-DVB extremely increased with increase in the adsorbent's pore size (molecular mass exclusions; M(lim)) from 2 x 10(3) to 1 x 10(4), but decreased with increase in the buffer's ionic strength (mu) to 0.2 or stronger. As a result, when the DMP-DVB (80:20) with M(lim) 2 x 10(3) was used as adsorbent by a column method at pH, 7.2 and mu = 0.17, it only selectively removed DNA from a BSA solution, including 1000 microg/ml of BSA and 10 microg/ml of DNA. The adsorbent decreased the concentration of DNA in the BSA solution to < 10 ng/ml, and the recovery rate of BSA was more 98%. PMID- 15043262 TI - Differences in preparative loadability between the charged and uncharged forms of ionizable compounds. AB - Ionizable compounds experience a drastic difference in preparative loadability as a function of pH. It can be shown that the preparative loadability of a compound in the ionic form is by a factor of 20 or more inferior to the loadability of the same compound in the unionized form. In this paper, we demonstrate the reason for this behavior, and show practical applications of the principle. PMID- 15043263 TI - Testing of "special base" columns in reversed-phase liquid chromatography. A rational approach considering solvent effects. AB - A methodology for building a chromatographic test aiming at characterizing special base stationary phases was described. Instead of choosing its conditions a priori, a "full" comprehensive test based on extended running conditions was performed on a 12 column set. The conditions were carefully chosen from their ability to take into account the solvent and the pH effects. Principal component analysis (PCA) has been combined to hierarchical cluster analysis both to provide interpreted classifications and to reduce drastically the test itself by eliminating redundant information. The final reduced test can be considered optimal because the minimized set of test conditions allows to provide as much information as in the initial full test. PMID- 15043264 TI - Rapid method for evaluating reversed-phase high-performance liquid chromatography column stability. AB - A procedure is presented for the rapid evaluation of HPLC stationary phase stability at pH 8.4 or 10.1 using a temperature of 60 degrees C. Mobile phase (MeOH-0.1 mol l(-1) aqueous NaHCO3, 50:50, v/v) is continuously passed through the column with periodic injections of a test solution until the several chromatographic parameters of the resulting chromatograms are degraded. The tests were applied to several commercial and laboratory-made stationary phases. After degradation two of these phases, one commercial and one laboratory-made, were examined by elemental analysis and scanning electron microscopy to elucidate the degradation process. PMID- 15043265 TI - Chromatographic and column stability at pH 7 of a C18 dimethylurea polar stationary phase. AB - Chromatographic evaluations of a C18 dimethylurea phase in 150 mm x 3.9 mm HPLC columns were performed using the Tanaka and Engelhardt test mixtures. The applicability of the new C18 dimethylurea phase was also evaluated with a mixture of some herbicides and their metabolites. An artificial aging procedure was also performed by passing a potassium phosphate mobile phase buffered at pH 7.0 through C18 50 mm x 3.9 mm dimethylurea columns. The column stability was evaluated by means of the chromatographic parameters obtained for the separation of some compounds from the Neue test mixture, using apolar, polar and highly basic analytes. PMID- 15043266 TI - Evaluation of packed capillary liquid chromatography columns and comparison with conventional-size columns. AB - Apart from extracolumn effects peak dispersion in liquid chromatographic columns is caused by the column inlet, the packed bed, and the column outlet. A strategy applicable for independent evaluation of the individual sources of column band broadening was developed on the basis of the linear extrapolation method (LEM). This method was applied to compare the performance of packed capillary LC columns from various commercial suppliers with conventional-size columns. The columns differed widely in their performance with respect to peak shapes and widths for standard substances. The capillary columns were found well packed, but in some cases overall performance would benefit from improving the design of the area between the packed bed and the connecting capillaries, containing frits as well as dead volumes. PMID- 15043267 TI - Model column structure for the analysis of the flow and band-broadening characteristics of silica monoliths. AB - We report on the use of commercial computational fluid dynamics software to study the band broadening in a perfectly ordered three-dimensional model structure, the so-called tetrahedral skeleton column (TSC), selected for its close geometrical resemblance to the specific pore network topology of silica monoliths. Van Deemter plots are presented for the case of a species flow through a non-porous skeleton and for the case of a retained component (k' = 1) in a porous skeleton (mesopore porosity epsilon = 0.6 in both cases). Using the flow domain as the characteristic scaling dimension, the TSC model yields reduced plate heights as small as h(min) = 0.8 and separation impedances as small as Emin = 120 for a retained component with k' = 1. The very small reduced plate heights for the TSC model can without any doubt largely be attributed to the perfect homogeneity of the considered model structure: the B and C terms are similar to those obtained in real silica monoliths with similar external porosity, whereas the A term is significantly (about a factor of 10) smaller. The present study hence suggests that further experimental work to obtain more homogeneous silica networks could yield large gains in reduced plate height and separation impedance. Comparing the three-dimensional TSC model with a 2D array of cylinders, it was found that the use of the domain size as the characteristic dimension in the reduced plate height expression is much more appropriate than the use of the skeleton size, hence validating earlier approaches adopted in the literature. PMID- 15043268 TI - Potential of silica monolithic columns in peptide separations. AB - The objective of the work described here was to evaluate the efficacy of silica monolith supports in high-speed reversed-phase liquid chromatography (RPLC) of peptides. This was done using a commercial Chromolith column with an octadecylsilane stationary phase and a tryptic digest of cytochrome c. Columns (100 mm x 4.6 mm) were operated at mobile phase velocities ranging from 1 ml/min (2.0 mm/s) to 10 ml/min (25 mm/s). There was little noticeable change over this flow rate range in either resolution, peak elution volume, or analyte concentration in collected fractions. It was concluded that capillary columns in this silica monolith format would be particularly valuable in peptide separations for proteomics. There was, however, a small, but perceptible contamination of peaks at high mobile phase velocity with earlier eluting analytes. Based on the fact that peak shape did not change at high mobile phase velocity, it is suggested that this phenomena might be due to the presence of peptide conformers in structural equilibrium on the sorbent surface. When elution rate exceeds the rate of conformer interchange, conformers could elute as broadened or even separate peaks. PMID- 15043269 TI - Comparison of styrene-divinylbenzene-based monoliths and Vydac nano-liquid chromatography columns for protein analysis. AB - This report details the fabrication of polystyrene-based nano-LC monolithic columns for the separation of protein molecules. The report focuses on the practical advantages of monolithic columns when compared to conventional packed columns. Capillary columns were made to 100 and 50 microm i.d. and used to analyse a mixture of proteins, these separations were then compared with a conventional protein phase under the same conditions. A second functionalised monolithic polystyrene-based column was also manufactured and compared for the analysis of proteins, under the same analytical conditions, to the standard non functionalised bare polystyrene monolith. Nano-LC polystyrene monolithic columns were found to be advantageous to conventional phases for the analysis of protein molecules, with a one-step fabrication process, faster analysis times, lower limits of detection hence higher sensitivity. PMID- 15043270 TI - Characterization of some physical and chromatographic properties of monolithic poly(styrene-co-divinylbenzene) columns. AB - Monolithic capillary columns were prepared by copolymerization of styrene and divinylbenzene inside a 200 microm i.d. fused silica capillary using a mixture of tetrahydrofuran and decanol as porogen. Important chromatographic features of the synthesized columns were characterized and critically compared to the properties of columns packed with micropellicular, octadecylated poly(styrene-co divinylbenzene) (PS-DVB-C18) particles. The permeability of a 60 mm long monolithic column was slightly higher than that of an equally dimensioned column packed with PS-DVB-C18 beads and was invariant up to at least 250 bar column inlet pressure, indicating the high-pressure stability of the monolithic columns. Interestingly, monolithic columns showed a 3.6 times better separation efficiency for oligonucleotides than granular columns. To study differences of the molecular diffusion processes between granular and monolithic columns, Van Deemter plots were measured. Due to the favorable pore structure of monolithic columns all kind of diffusional band broadening was reduced two to five times. Using inverse size exclusion chromatography a total porosity of 70% was determined, which consisted of internodule porosity (20%) and internal porosity (50%). The observed fast mass transfer and the resulting high separation efficiency suggested that the surface of the monolithic stationary phase is rather rough and does not feature real pores accessible to macromolecular analytes such as polypeptides or oligonucleotides. The maximum analytical loading capacity of monolithic columns for oligonucleotides was found to be in the region of 500 fmol, which compared well to the loading capacity of the granular columns. Batch-to-batch reproducibility proved to be better with granular stationary phases compared to monolithic stationary phase, in which each column bed is the result of a unique column preparation process. PMID- 15043271 TI - Titanized silica-based stationary phases prepared with thermally and microwave immobilized poly(methyloctylsiloxane). AB - Silica supports having their surface modified with titanium oxide were prepared and coated with poly(methyloctylsiloxane) (PMOS). Subsequently, immobilization of the polysiloxane was induced by thermal treatment or microwave radiation. The thermal treatment was carried out for different times (4, 8, 16 and 24 h) at temperatures ranging between 100 and 220 degrees C. For PMOS immobilization by microwave radiation, 452, 520 and 586 W power levels and exposure times of 5, 15 and 30 min were used. After extraction of non-immobilized polymer, the chromatographic properties of the phases were evaluated. The phase immobilized at 120 degrees C for 8 h presented the best chromatographic parameters, suggesting that the quantity of acidic hydroxyl groups on the support surface was reduced, resulting in fewer undesirable interactions of a basic solute with the silanols not removed or covered on the support surface. PMID- 15043272 TI - Self-immobilization and/or thermal treatment for preparing silica poly(methyloctylsiloxane) stationary phases. AB - Batches of poly(methyloctylsiloxane) (PMOS)-loaded silica were prepared by the deposition of PMOS, into the pores of HPLC silica. Portions of PMOS-loaded silica were allowed to remain at ambient temperature, without further treatment for 2, 9, 20, 31, 51, 105 and 184 days after preparation to undergo self-immobilization (irreversible adsorption of a layer of polymer on silica at ambient temperature in the absence of initiators). Other portions were subjected to a thermal treatment (100 degrees C for 4h) after 1, 2, 5, 7, 9, 15, 20, 25, 70, 111 and 184 days. Self-immobilized and thermally treated samples were characterized by % C, 29Si cross-polarization magic angle spinning (CP/MAS) NMR spectroscopy and reversed-phase column performance. The results show that thermal immobilization accelerates the distribution and rearrangement of the polymer on the silica surface. However, from the time that a monolayer has been formed by self immobilization (approximately 100 days for PMOS on Kromasil silica), the thermal treatment does not alter this configuration and, thus, does not change the resulting chromatographic parameters. PMID- 15043273 TI - Microwave-immobilized polybutadiene stationary phase for reversed-phase high performance liquid chromatography. AB - Polybutadiene (PBD) has been immobilized on high-performance liquid chromatography (HPLC) silica by microwave radiation at various power levels (52 663 W) and actuation times (3-60 min). Columns prepared from these reversed-phase HPLC materials, as well as from similar non-irradiated materials, were tested with standard sample mixtures and characterized by elemental analysis (%C) and infrared spectroscopy. A microwave irradiation of 20 min at 663 W gives a layer of immobilized PBD that presented good performance. Longer irradiation times give thicker immobilized layers having less favorable chromatographic properties. PMID- 15043274 TI - Influence of air on polybutadiene used in the preparation of stationary phases for high-performance liquid chromatography. AB - A 100 ml bottle of polybutadiene, PBD, was repeatedly exposed to air over a period of 6 months. Samples were taken at time zero (PBD-0), after 3 months (PBD 3) and 6 months (PBD-6). These samples were sorbed onto HPLC silica by an open air solution-evaporation procedure, which involved exposure to the atmosphere for 6 days. Portions of the three sets of samples were used to compare self immobilization and the effects of 100 degrees C thermal treatments in air or nitrogen on HPLC performance of the resulting phases. It is concluded that self immobilization is enhanced by prior exposure of sorbed PBD to air and subsequent heating at 100 degrees C further enhances column performance. The best performance (10(5) plates m(-1)) resulted from 4 h heating of PBD-6 material in nitrogen. PMID- 15043275 TI - Novel surface-modification techniques for polymer-based separation media. Stimulus-responsive phenomena based on double polymeric selectors. AB - A pair of polymeric selectors potentially responding to stimulation was introduced onto monosized porous polymer particles to be evaluated as a packing material for HPLC. Possible complexes formed between polyacrylamide (PAAm) and poly(methacrylic acid) (PMAA) were utilized as stimulus responsive polymeric selectors. Uniformly sized base polymer particle was prepared by multi-step swelling and polymerization method, while the introduction of PAAm and PMAA was done by newly invented modification technique. In this technique, a solvent in which both acrylamide (AAm) and methacrylic acid (MAA) monomers are soluble, but PAAm and PMAA are insoluble, was utilized as a modification medium. The polymer particle doubly modified with PAAm and PMAA was utilized as packing material for HPLC and the stimulus responses were evaluated by changing temperature or pH to check change of the slope of a Van't Hoff plot. By using water as a mobile phase, the expected inflection point of the Van't Hoff plot was observed at upper critical solution temperature (UCST) of the polymer complexes and the temperature responsive ability was observed. Moreover, pH responsive ability was studied by using buffer of either pH 4 or 10 as mobile phase. Slope of the plot was changed in buffer of pH 4, but no change of slope was observed in the buffer of pH 10. PMID- 15043276 TI - Temperature- and pH-responsive aminopropyl-silica ion-exchange columns grafted with copolymers of N-isopropylacrylamide. AB - We have designed copolymers of N-isopropylacrylamide, environmentally-responsive polymers, which respond to temperature and other external stimuli. In this study, we designed and synthesized copolymers that introduced ion-exchange groups. These copolymers responded to the temperature and the pH, and the copolymer-grafted aminopropyl silica beads were used as HPLC packing materials. This stationary phase altered the properties from hydrophilic to hydrophobic and from charge to non-charge by temperature and pH changes. We studied the separations of organic acids and phenylthiohydantoin-amino acids using environmentally-responsive chromatography, and confirmed the effects of the ion-exchange groups. The elution behaviors of these samples were controlled by the temperature changes without organic solvents in the mobile phase. It was confirmed that the interactions between the solute and stationary phase could be freely controlled by the temperature and the pH. Environmentally-responsive chromatography is expected to be applicable to the separation of pharmaceuticals and biomolecules, such as peptides, proteins and nucleic acids. PMID- 15043277 TI - Effect of high-temperature on high-performance liquid chromatography column stability and performance under temperature-programmed conditions. AB - Six commercially available analytical (4.1 or 4.6 mm i.d.) columns were evaluated under temperature-programmed high-temperature liquid chromatography (HTLC) conditions to access their stability and performance at extreme temperatures. Seven components consisting of acidic, basic and neutral compounds were analyzed under temperature-programmed conditions and solvent gradient conditions using three different mobile phase compositions (acidic, basic and neutral). Each column was checked with a two-component test mix at various stages of the evaluation to look for signs of stationary phase collapse. Three zirconia based stationary phases studied exhibited column bleed under temperature-programmed conditions. The other three columns, a polydentate silica column, a polystyrene divinylbenzene (PS-DVB) polymeric column, and a graphitic carbon column performed well with no evidence of stationary phase degradation. The R.S.D. for the retention times and efficiencies were less than 10% for most conditions, and not more than 15% during the course of the evaluation for each column. The polydentate silica stationary phase was temperature programmed to 100 degrees C, the PS-DVB stationary phase was temperature programmed up to 150 degrees C, and the graphitic carbon column was used with temperature programming up to 200 degrees C. Comparable peak capacities and similar retention behaviors were observed under solvent gradient and temperature-programmed conditions. Temperature programming with dynamic mobile phase preheating can replace solvent gradient analysis without a loss of peak capacity when used with 4.1 or 4.6 mm columns. PMID- 15043278 TI - Suppression of deleterious effects of free silanols in liquid chromatography by imidazolium tetrafluoroborate ionic liquids. AB - Silica-based stationary phases are commonly used in liquid chromatography, but their surface acidity causes known problems, especially when separating basic compounds. Deleterious effects of free silanols are not fully removed by standard prevention procedures consisting in adding alkylamines or other amino quenchers to the eluents. We found that ionic liquids of the imidazolium tetrafluoroborate class, added to mobile phases at concentrations of 0.5-1.5% (v/v), blocked silanols and provided excellent thin-layer chromatographic separations of strongly basic drugs which were otherwise not eluted, even with neat acetonitrile as the mobile phase. The silanol suppressing potency of imidazolium tetrafluoroborates was demonstrated to markedly exceed that of the standard mobile phase additives, like triethylamine, dimethyloctylamine and ammonia. The proposed new mobile phase additives were also demonstrated to provide reliable lipophilicity parameters of base drug analytes as determined by gradient mode of high-performance liquid chromatography. By applying the readily available and environmentally friendly imidazolium tetrafluoroborate ionic liquids, simple and efficient means of improvement of liquid chromatographic analysis of organic bases were elaborated. PMID- 15043279 TI - Effects of ions and detergents in drug partition chromatography on liposomes. AB - We have determined drug partitioning into phospholipid bilayers by immobilized liposome chromatography (ILC). Electrostatic effects on the drug partitioning were observed on neutral bilayers at low ionic strength. The size of the counterions affected the partitioning. When liposomes were supplemented with ionic detergents the partitioning of charged drugs was strongly affected, allowing complete separation of drugs of different charges which showed similar retention on neutral bilayers. Partial separation was obtained on bilayers containing fatty acid. Detergent ions or fatty acid inserted into phospholipid bilayers affected the partitioning of drugs much more than did free ions or phospholipid head group charges. PMID- 15043280 TI - Micellar versus hydro-organic mobile phases for retention-hydrophobicity relationship studies with ionizable diuretics and an anionic surfactant. AB - Logarithm of retention factors (log k) of a group of 14 ionizable diuretics were correlated with the molecular (log P o/w) and apparent (log P(app)) octanol-water partition coefficients. The compounds were chromatographed using aqueous-organic (reversed-phase liquid chromatography, RPLC) and micellar-organic mobile phases (micellar liquid chromatography, MLC) with the anionic surfactant sodium dodecyl sulfate (SDS), in the pH range 3-7, and a conventional octadecylsilane column. Acetonitrile was used as the organic modifier in both modes. The quality of the correlations obtained for log P(app) at varying ionization degree confirms that this correction is required in the aqueous-organic mixtures. The correlation is less improved with SDS micellar media because the acid-base equilibriums are shifted towards higher pH values for acidic compounds. In micellar chromatography, an electrostatic interaction with charged solutes is added to hydrophobic forces; consequently, different correlations should be established for neutral and acidic compounds, and for basic compounds. Correlations between log k and the isocratic descriptors log k(w), log k(wm) (extrapolated retention to pure water in the aqueous-organic and micellar-organic systems, respectively), and psi0 (extrapolated mobile phase composition giving a k = 1 retention factor or twice the dead time), and between these descriptors and log P(app) were also satisfactory, although poorer than those between log k and log P(app) due to the extrapolation. The study shows that, in the particular case of the ionizable diuretics studied, classical RPLC gives better results than MLC with SDS in the retention hydrophobicity correlations. PMID- 15043281 TI - Ultrapure water for liquid chromatography-mass spectrometry studies. AB - Improvements in trace enrichment techniques combined with the sensitivity of mass spectrometry offer enhanced opportunities to analyze ever lower concentrations of drugs, metabolites, pesticides or environmental pollutants. To perform HPLC and liquid chromatography-mass spectrometry (LC-MS) analyses under optimum conditions, the water used for mobile phase preparation needs to be highly purified and delivered on demand. Indeed, both UV photodiode array detection and MS detection methods are sensitive to organic contaminants (total organic carbon, TOC), and the water quality has a direct impact on the achievable detection limits. The benefits of UV photooxidation on TOC reduction for LC-MS studies were highlighted using electrospray ionization MS detection by comparing HPLC-grade bottled water, freshly produced UV185/254-treated water, and freshly produced non UV-treated water. PMID- 15043282 TI - The rectogenital septum: morphology, function, and clinical relevance. AB - PURPOSE: The rectogenital septum (known in clinical literature as Denonvilliers' fascia) forms an incomplete partition between the rectum and the urogenital organs in both men and women. It is composed of collagenous and elastic fibers and smooth muscle cells intermingled with nerve fibers emerging from the autonomic inferior hypogastric plexus. The aim of this study was to investigate the fetal development of the rectogenital septum, and the origin and innervation of the longitudinal smooth muscle cells within the septum, as well as to consider possible effects on function of operations that compromise the integrity of these structures. METHODS: Macroscopic dissections on embalmed human pelves and plastination histology of 40 fetal and newborn pelvic specimens were performed. By means of conventional and immunohistochemical staining methods using monoclonal and polyclonal antibodies for tissue analysis and neuronal labeling, the motor and sensory innervation of the longitudinal muscle bundles within the septum was defined. RESULTS: The rectogenital septum is formed by a local condensation of mesenchymal connective tissue in the early fetal period. The longitudinal muscle bundles could be traced back to the longitudinal layer of the rectal wall, and, using the septum as a guiding structure, it was possible to identify autonomic nerve fibers and ganglion cells innervating the muscle cells and crossing the midline without detectable gender differences. CONCLUSIONS: Because of a coinnervation of the rectal muscle layers and the adjacent longitudinal muscle fibers of the septum, a functional correlation between the two structures during defecation is postulated. On the basis of these findings, a safer dissection of the anterior rectal wall during rectal resection is postulated, thus limiting functional disturbance and preventing neural damage. PMID- 15043283 TI - Gene expression profiling in lymph node-positive and lymph node-negative colorectal cancer. AB - PURPOSE: To identify the genes involved in the carcinogenesis and progression of colorectal cancer, we analyzed the gene-expression profiles of colorectal cancer cells from 12 tumors with corresponding noncancerous colonic epithelia using a cDNA microarray representing 4,08 genes. METHODS: We classified both samples and genes by using a two-way clustering analysis and identified genes that were differentially expressed in the cancerous and noncancerous tissues. Genes associated with lymph node metastasis were identified by means of the supervised learning technique. RESULTS: Differentially expressed genes (77 up-regulated and 45 down-regulated genes) were identified in more than 75 percent of the tumors. The functional categories of these genes belonged to signal transduction (19 percent), metabolism (17 percent), cell structure/motility (14 percent), cell cycle (13 percent), and gene protein expression (13 percent). The gene expression pattern of reverse transcriptase polymerase chain reaction (RT-PCR) results from randomly selected genes shows a pattern similar to that of cDNA microarray. Moreover, the gene expression patterns observed were similar to those reported previously, suggesting rare racial differences. Sixty genes possibly associated with lymph node metastasis in colorectal cancer were selected on the basis of clinicopathological data obtained by performing signal-to-noise calculations. "Leave-one-out" cross-validation testing correctly classified 10 of 12 patients (83.3 percent) as having colorectal cancer with lymph node metastasis vs. those without metastasis. CONCLUSIONS: These results provide not only a new molecular basis for understanding the biologic properties of colorectal cancer, including lymph node metastasis, but also provide a resource for future development of therapeutic targets and diagnostic markers for colorectal cancer. PMID- 15043284 TI - Acceptance of and attitude toward genetic testing for hereditary nonpolyposis colorectal cancer: a comparison of participants and nonparticipants in genetic counseling. AB - PURPOSE: Data on the actual uptake of genetic testing for hereditary nonpolyposis colorectal cancer (HNPCC) in a clinical sample is still inconclusive. The present study aimed to define the actual uptake of genetic counseling and testing offered to an unselected sample of 140 patients with colorectal cancer, fulfilling clinical criteria (Amsterdam or Bethesda) suggestive of HNPCC, and to identify demographic and psychosocial factors associated with the decision to participate in counseling. METHODS: Cross-sectional survey. Eligible subjects had been consecutively enrolled in a regional tumor registry between 1994 and 1998, and were invited to attend an information session for HNPCC at the time genetic testing for HNPCC became available. Participants and nonparticipants in the information session completed a short questionnaire. RESULTS: The actual uptake rate of the information session in this sample was 26 percent. Participants and nonparticipants were comparable with regard to clinical criteria suggestive of HNPCC, awareness of the potential hereditary predisposition, and previous history of cancer in the family. Some 60 percent of participants experienced pronounced distress related to their potential inheritance of the disorder, compared to 35 percent among nonparticipants. Distress reached a clinically significant level in 28 percent of participants. Restricted communication within the family was observed frequently. Irrespective of groups, a positive attitude toward obtaining a gene test result predominated. CONCLUSIONS: Results suggest that expressed intention and attitude toward genetic testing do not reliably predict actual uptake of counseling or testing. Thorough interdisciplinary counseling should be provided to every patient with clinical criteria suggestive of HNPCC. The considerable distress related to the hereditary disorder should be adequately addressed, as should be communication issues. PMID- 15043285 TI - Neuroendocrine carcinomas of the colon and rectum. AB - PURPOSE: This study was designed to review experience with neuroendocrine carcinomas of the colon and rectum at a single institution, with emphasis on the pathology and clinical characteristics of this uncommon malignancy. METHODS: A study group of patients was identified from a prospective colorectal service database. Pathology was reviewed and neuroendocrine tumors were classified by a single pathologist. Medical records were retrospectively reviewed. RESULTS: From March 1975 to September 1998, 38 patients with neuroendocrine carcinomas were identified from the colorectal service database comprising 6495 patients (0.6 percent). These neuroendocrine carcinomas did not include carcinoid tumors. Average patient age was 57 years (range, 29-86 years). There were 17 males (44.7 percent) and 21 females (55.3 percent). Tumors were located as follows: 17 colon, 14 rectum, 6 anal canal, and 1 appendix. The diagnosis of neuroendocrine carcinoma was suggested preoperatively from tissue biopsy in 59.3 percent (16/27) of patients evaluable. Pathology was reviewed and tumors were categorized as small cell carcinoma (n = 22) or large cell neuroendocrine carcinoma (n = 16). Most tumors (20/25 evaluable, 80 percent) stained positive by means of immunohistochemistry for neuroendocrine markers, including chromogranin (18/19), synaptophysin (10/15), and/or neuron-specific enolase (14/15). Metastatic disease was detected at the time of diagnosis in 69.4 percent of the patients (25/36). Tumors were advanced at the time of diagnosis, with American Joint Committee on Cancer (AJCC) Stage I (n = 6), Stage III (n = 7), and Stage IV (n = 25) tumors. As a group, these tumors had a poor prognosis, with a median survival of 10.4 months. One-year, two-year, and three-year survival was 46 percent, 26 percent, and 13 percent, respectively. There was no significant difference in survival based on pathologic subtypes. Median follow-up time was 9.4 months (range, 0.6 263.7 months). CONCLUSIONS: Neuroendocrine carcinomas of the colon and rectum are uncommon, comprising less than 1 percent of colon and rectal cancers. Pathologically, these tumors are poorly differentiated carcinomas with distinctive cytoarchitectural features and are often immunoreactive for markers of neuroendocrine differentiation. The prognosis for high-grade neuroendocrine carcinomas is poor, as most patients have metastatic disease at the time of diagnosis. PMID- 15043286 TI - Preoperative radiotherapy combined with intraoperative radiotherapy improve results of total mesorectal excision in patients with T3 rectal cancer. AB - PURPOSE: The survival advantage of preoperative radiotherapy in patients with rectal cancer is still a matter of debate, because its incremental benefit in the total mesorectal excision setting is unclear. This study was designed to evaluate early and long-term results of preoperative radiotherapy plus intraoperative radiotherapy in a homogeneous population of T3 middle and lower rectal cancer patients submitted to total mesorectal excision. METHODS: A series of 113 patients with middle and lower T3 rectal cancer consecutively submitted to total mesorectal excision at a single surgical unit from 1991 to 1997 were divided into two groups according to type of neoadjuvant treatment: preoperative radiotherapy (38 Gy) plus intraoperative radiotherapy (10 Gy; n = 69), and no preoperative treatment (total mesorectal excision; n = 44). Standard statistical analyses were used to evaluate early (downstaging, intraoperative factors, hospital morbidity, and mortality rates) and long-term results (recurrence and survival). RESULTS: Overall, 68.2 percent of patients were downstaged by the preoperative regimens (T0 specimens in 3 cases). Postoperative complications were comparable in the two groups. Five-year, disease-specific survival was 81.4 and 58.1 percent in preoperative radiotherapy plus intraoperative radiotherapy group and total mesorectal excision group, respectively (P = 0.052). Corresponding figures for disease-free survival were 73.1 and 57.2 percent in the two groups, respectively (P = 0.096). The rates of local recurrence at five years were 6.6 and 23.2 percent in preoperative radiotherapy plus intraoperative radiotherapy and total mesorectal excision groups, respectively (P = 0.017). CONCLUSIONS: Preoperative radiotherapy plus intraoperative radiotherapy associated with total mesorectal excision reduce local recurrence rate and improve survival in T3 rectal cancer compared with total mesorectal excision alone. PMID- 15043287 TI - Outcome of patients with ischemic colitis: review of fifty-three cases. AB - PURPOSE: Ischemic colitis is a disease of elderly patients and includes a wide clinical spectrum ranging from mild to severe forms. Some patients may develop complications. Management of this disorder depends on disease severity. Our aim was to review the clinical characteristics of patients diagnosed of ischemic colitis and analyze predictive factors of poor prognosis. METHODS: This study is a retrospective analysis of 53 cases of ischemic colitis (33 men, 20 women), 35 with moderate and 18 with severe forms, respectively. Clinical characteristics, diagnostic procedures, segment of colon involved and long-term evolution after discharge were analyzed. RESULTS: Hypertension (51 percent) was the main risk factor associated with ischemic colitis. Clinical presentation did not differ between groups, except for peritonitis which was present only in the severe group. Colonoscopy and histologic studies were the most used diagnostic procedures (90 percent). Peripheral vasculopathy (P < 0.01) and right colonic involvement (P < 0.001) were risk factors for severe outcome. Five patients died during admission. Among these, the right colon was affected in four (80 percent). No patient in either group developed chronic ischemic colitis during follow-up. CONCLUSION: Ischemic colitis usually runs a benign course after acute colonic insult. Peripheral vasculopathy and right colonic involvement are associated with severe forms of ischemic colitis. PMID- 15043288 TI - Repair of paracolostomy hernias with a prosthetic mesh in the intraperitoneal onlay position: modified Sugarbaker technique. AB - PURPOSE: The disappointing outcome of local fascial repair and stoma relocation in parastomal hernias has stimulated a variety of new techniques that use a prosthetic mesh for herniorrhaphy. Many of these procedures either carry the risk of mesh contamination or allow only a local repair. We established a method that allows both an aseptic operation and the management of concurrent incisional hernias. METHODS: In a retrospective study we evaluated all patients who had undergone operation for a paracolostomy hernia with an expanded polytetrafluoroethylene (PTFE) mesh in the intraperitoneal onlay position in our Department of General Surgery from 1994 until 2002. Twenty patients with large paracolostomy hernias and 10 additional ventral hernias, mostly large incisional or recurrent incisional hernias, were identified. We combined the Sugarbaker and the Rives-Stoppa techniques by covering the defects with an ePTFE mesh after laparotomy and fixing the mesh with traction sutures. RESULTS: Postoperatively, there was no incidence of mesh infection. After a mean follow-up of 3 1/2 years we found three recurrences of paracolostomy hernias and two recurrences of incisional hernias. Another two hernias emerged in the previously intact midline. All these hernias were small, without tendency to enlargement, and did not warrant reoperation. CONCLUSIONS: Patients with symptomatic paracolostomy hernias or a combination of abdominal wall defects should properly be managed surgically with an intraperitoneally placed mesh that covers all hernias. Our results, with a recurrence rate of only 15 percent for the parastomal site and 20 percent for combined defects, support this approach. PMID- 15043289 TI - Neural network analysis of anal sphincter repair. AB - PURPOSE: Prediction of success after anterior sphincter repair for incontinence is difficult. Standard multivariate analysis techniques have only 75 to 80 percent accuracy. Artificial intelligence, including artificial neural networks, has been used in the analysis of complex clinical data and has proved to be successful in predicting the outcome of other surgical procedures. Using a neural network algorithm, we have assessed the probability of success after anterior sphincter repair. METHODS: Prospective anorectal physiology data of 72 patients undergoing anterior sphincter repair was collected between 1995 and 1999. Complete data sets of 75 percent of the series were used to train an artificial neural network; the remaining 25 percent were used for data validation. The output was continence grading, ranging from 0 to 4 (worse to continent). RESULTS: The outcome at 3, 6, and 12 months postoperatively was obtained and assessed. The best correlation between actual data value and artificial neural network value was found at 12 months (r = 0.931; P = 0.0001). Clear correlations also were found at three months (r = 0.898; P = 0.0001) and six months (r = 0.742; P = 0.002). Results of applying a net to details excluding pudendal nerve latency were poor. CONCLUSIONS: Artificial neural networks are more accurate (93 percent correlation) than standard statistics (75 percent) when applied to the prediction of outcome after anterior sphincter repair. This assessment also confirms the usefulness of pudendal latency in the prediction of anterior sphincter repair outcome. The results obtained highlight the obvious usefulness of artificial neural networks, which could now be used in a prospective evaluation for application of the technique. PMID- 15043290 TI - Anal sphincter morphology in patients with full-thickness rectal prolapse. AB - PURPOSE: The aim of this study was to assess the morphologic change of the anal canal in patients with rectal prolapse. METHODS: The endoanal ultrasound scans of 18 patients with rectal prolapse were compared with those of 23 asymptomatic controls. The thickness and area of the internal anal sphincter and submucosa were measured at three levels. RESULTS: Qualitatively, patients with rectal prolapse showed a characteristic elliptical morphology in the anal canal with anterior/posterior submucosal distortion accounting for most of the change. Quantitatively, internal anal sphincter (IAS) and submucosa (SM) thicknesses and area were greater in all quadrants of the anal canal (especially upper) in patients with rectal prolapse compared with controls. There was statistical evidence (in a regression model) of a relationship between increases in all measured variables and the finding of rectal prolapse. CONCLUSIONS: The cause of sphincter distortion in rectal prolapse is unknown but may be a response to increased mechanical stress placed on the sphincter from the prolapse or an abnormal response by the sphincter complex to the prolapse. Patients found to have this feature on endoanal ultrasound should undergo clinical examination and defecography to look for rectal wall abnormalities. PMID- 15043291 TI - New surgical options for fecal incontinence in patients with imperforate anus. AB - INTRODUCTION: Anorectal malformations are among the various etiologic factors causing fecal incontinence. Patients with imperforate anus are difficult to treat, specifically those with high lesions. The artificial bowel sphincter and electrically stimulated gracilis neosphincter are two relatively new techniques that have been used for the treatment of patients with severe refractory fecal incontinence. The aim of this study was to evaluate the results of these technologies in the treatment of patients with chronic fecal incontinence due to imperforate anus. METHODS: All patients with imperforate anus who had fecal incontinence and underwent either the artificial bowel sphincter procedure or the gracilis neosphincter procedure between February 1995 and December 2000 were evaluated. Preoperative and postoperative incontinence score (Cleveland Clinic Florida Incontinence Score; 0 = perfect continence; 20 = complete incontinence), quality of life, (Fecal Incontinence Quality of Life Scale, 29 items forming 4 scales), and manometric sphincter pressure results were compared. RESULTS: Eleven patients had artificial bowel sphincter and five had the gracilis neosphincter (one nonstimulated) procedure. There were 11 males and 5 females of a mean age of 25.3 (range, 15-45) years. The mean follow-up time was 1.7 years (5 months to 5.7 years). Eight (50 percent) complications occurred in six patients, including three with fecal impaction (all artificial bowel sphincter), three with device migration (two gracilis neosphincter, one artificial bowel sphincter), and two patients with concomitant wound infection (one gracilis neosphincter, one artificial bowel sphincter); no patients had the devices explanted. Fourteen patients had manometric data (10 artificial bowel sphincter and 4 gracilis neosphincter) available. The overall incontinence score decreased from a preoperative mean of 18.5 to a postoperative mean of 7.5 in the artificial bowel sphincter group (P < 0.01) and from 17.4 to 9.4 in the gracilis neosphincter group (P = 0.06). All four Fecal Incontinence Quality of Life scales increased in both the artificial bowel sphincter (lifestyle and depression/self-perception, P = 0.02; coping/behavior and embarrassment, P = 0.03) and the gracilis neosphincter (lifestyle and coping, P = 0.06; depression and embarrassment, P = 0.05) patients. As well, the mean resting and squeeze pressures increased with both techniques (artificial bowel sphincter: P = 0.008 and P = 0.02, respectively; gracilis neosphincter: P = 0.4 and P = 0.1, respectively). All results were statistically significant in the artificial bowel sphincter group. CONCLUSIONS: Artificial bowel sphincter and gracilis neosphincter are efficient methods to treat patients with imperforate anus. These techniques should be considered for patients with imperforate anus and severe fecal incontinence. PMID- 15043292 TI - Effect of neurotensin on colorectal motor and sensory function in humans. AB - PURPOSE: Animal studies have shown that neurotensin stimulates colonic motility, but little is known on the effect on rectocolonic function in humans. This study was designed to investigate the effect of neurotensin on rectal and colonic motor and sensory function and colonic reflexes in humans. METHODS: Motor and sensory function of the descending colon and rectum were studied in eight healthy volunteers (5 females; age range, 20-58 years) by using a dual, computerized, rectocolonic, barostat assembly. Measurements were performed during placebo and neurotensin infusion (5 pmol/kg per minute), respectively. Compliance and reflex mechanisms were assessed in both rectum and descending colon. Symptom perception (urge and pain) was scored using Visual Analog Scales (0-10 cm). RESULTS: Neurotensin significantly (P < 0.05) increased rectal compliance (from 9 +/- 1.1 to 10.1 +/- 1.1 ml/mmHg) and colonic compliance (from 7.9 +/- 0.4 to 9 +/- 0.7 ml/mmHg) during stepwise distensions. Intensity of urge and pain perception during rectal distension was increased by neurotensin (P < 0.05). Sensations during colonic distensions were not altered by neurotensin. For colonic reflexes, during rectal distension the colonic bag volume increased significantly (P < 0.05), whereas during colonic distension rectal bag volume decreased significantly (P < 0.05). Neurotensin delayed the occurrence of these reflexes: they were present at significantly higher pressures compared to placebo (P < 0.05). CONCLUSIONS: We demonstrated the presence of both a rectocolonic inhibitory and a colorectal stimulatory reflex in healthy volunteers. Neurotensin increases compliance and modulates rectal but not colonic sensitivity. Colorectal and rectocolonic reflexes are impaired during infusion of neurotensin. PMID- 15043293 TI - In vitro model for liposome-mediated adenomatous polyposis coli gene transfer in a duodenal model. AB - PURPOSE: Duodenal adenomas arise in more than 90 percent of patients with familial adenomatous polyposis (FAP). Management of severe duodenal disease remains difficult and controversial. This study investigates transfer of functional wild-type adenomatous polyposis coli (APC) gene under conditions of varying pH and bile concentrations into a somatic duodenal cancer cell line (HUTU 80) as a prelude to in vivo gene therapy in the management of severe duodenal disease. METHODS: In vitro transfection of human APC gene was performed on a human duodenal adenocarcinoma cell line (HUTU-80) by use of a liposomal vector. Different concentrations (5 percent and 10 percent) of human bile and varying pH (6 percent and 8 percent) were used during APC gene transfer to assess their effects on transfection efficiency. The effect of bile on the proliferation of HUTU-80 cells was evaluated by means of a colorimetric chemosensitivity assay with sulforhodamine B (SRB). The duration of APC transgene expression was analyzed by reverse transcription polymerase chain reaction (RT-PCR). RESULTS: Exogenous APC transgene was expressed in HUTU-80 cell line for 6 days after transfection. Differing pH did not affect APC gene transfer into the duodenal cell line with similar transgene expression to controls, but APC transfection efficiency was reduced semiquantitatively in the presence of bile. Coculturing with human bile (5 percent and 10 percent) did not affect the proliferation of HUTU-80. CONCLUSION: This study demonstrates transfer of APC gene into a duodenal epithelial cell line with prolonged transgene expression. Liposome-mediated APC gene transfer to the duodenum is feasible even in the presence of bile and varying pH, raising the potential of future gene therapy for this extremely difficult to treat condition. PMID- 15043295 TI - Limberg flap repair for pilonidal sinus disease. AB - PURPOSE: There are many methods described for the treatment of sacrococcygeal pilonidal sinus disease, and none of them has been accepted as an optimal modality. Plastic procedures have some advantages, such as short duration of hospitalization, quick healing time, low risk of wound infection, and lower recurrence rates. Our choice is Limberg flap repair; we present here our experience with this procedure. METHODS: From August 1998 to July 2000, 147 male patients were treated with Limberg flap repair under regional anesthesia in a soldier's hospital. RESULTS: No major anesthetic complication or wound infection developed. Three patients (2 percent) had a seroma (with negative culture) and six patients (4.1 percent) had partial wound detachment. Patients returned to full activity on the 10th to 25th postoperative day (mean, 18.8). Patients were followed from 1 to 40 (mean; 13.1) months. Seven patients (4.8 percent) had a recurrence. CONCLUSION: The Limberg flap procedure is an easy and effective technique. Patient comfort, quick healing time, early return to full activity, and low complication and recurrence rates are the important advantages of this procedure. PMID- 15043294 TI - Does serum CA19-9 play a practical role in the management of patients with colorectal cancer? AB - PURPOSE: CA19-9 is often used in combination with carcinoembryonic antigen to manage patients with colorectal cancer, even though there is insufficient evidence to support this use of CA19-9. Carcinoembryonic antigen, by contrast, has been regarded as a better indicator of poor prognosis and recurrence. The purpose of this study is to clarify whether CA19-9 is, in fact, a useful marker in the management of colorectal cancer patients by comparing it with carcinoembryonic antigen. METHODS: A retrospective investigation was done for a consecutive series of 155 patients with colorectal adenocarcinoma who underwent potentially curative surgery between 1995 and 1999. Excluded were patients with postoperative assays performed less than three times for either carcinoembryonic antigen or CA19-9 and those who had developed secondary cancers. Data from 118 patients were analyzed in terms of prediction of prognosis and detection of recurrences. RESULTS: The sensitivities of preoperative CA19-9 and carcinoembryonic antigen were 29.8 percent and 45.3 percent, respectively. In the univariate analysis of preoperative carcinoembryonic antigen and CA19-9 assays in 114 patients, high carcinoembryonic antigen level was significantly associated with poor prognosis (P = 0.0090) by log-rank test). We could not find a significant association between preoperative CA19-9 abnormality and survival (P = 0.12). Multivariate analysis of preoperative factors indicated significance in TNM stage (P = 0.0094) and tumor location (P = 0.036) but in neither carcinoembryonic antigen (P = 0.061) nor CA19-9 (P = 0.22). Among 40 patients with recurrences, postoperative elevations of tumor markers were seen in 19 cases for CA19-9 and in 37 for carcinoembryonic antigen throughout the follow-up periods. Sensitivity, specificity, positive predictive value, and negative predictive value were 0.48, 0.88, 0.68, and 0.77, respectively, for CA19-9, and 0.93, 0.88, 0.80, and 0.96, respectively, for carcinoembryonic antigen. In patients with recurrences, the initial postoperative elevation of tumor markers was seen earlier than the detection of recurrence in 68.4 percent of those with CA19-9 elevation and in 67.6 percent of those with carcinoembryonic antigen elevation. There was only one patient with recurrence who had CA19-9 elevation without carcinoembryonic antigen elevation, while 19 recurrent patients had carcinoembryonic antigen elevation without CA19-9 elevation. Multivariate analysis showed a significant risk of carcinoembryonic antigen elevation against recurrence with an odds ratio of 32.0 (P < 0.0001), in contrast to an insignificant association of CA19-9 elevation (P = 0.23). CONCLUSION: We could not find clinical significance to support the use of CA19-9 to predict the prognosis and detect recurrence of colorectal cancer. Because of this, we do not recommend routine use of CA19-9 in staging and surveillance of colorectal cancer patients. PMID- 15043297 TI - Ileal pouch perforation in pregnancy: report of a case and review of the literature. AB - Acute abdominal pain, especially in the presence of free intra-abdominal air, in a pregnant patient who has previously undergone ileal pouch-anal anastomosis may be a result of acute perforation of the pouch. The case of a 30-year-old multigravida with an uncomplicated pregnancy at 27 weeks gestation who was admitted to the perinatal ward for preterm labor is described. The patient's past medical history was significant for an ileal pouch-anal anastomosis for ulcerative colitis. Forty-eight hours after admission, she acutely developed generalized abdominal pain with associated dyspnea. A spiral CT of the chest to rule out pulmonary embolus revealed free intra-abdominal air. An emergency cesarean section was performed, which resulted in a viable female infant. The etiology for the free air was a perforation of the ileal pouch secondary to adhesions to the posterior part of the uterus. Perforation of the pouch has been reported secondary to a variety of factors. However, no cases of ileal pouch perforation in pregnancy have been reported. Adhesions that involve the pouch and an enlarged contracting uterus may result in perforation of the pouch. Awareness and knowledge of this potential complication may lead to earlier recognition and optimal management. PMID- 15043296 TI - Picoprep-3 is a superior colonoscopy preparation to Fleet: a randomized, controlled trial comparing the two bowel preparations. AB - PURPOSE: Bowel preparations for colonoscopy have to balance the demand for adequate cleansing action of the bowel and patient acceptability. There has been no study comparing Picoprep-3 (sodium picosulfate), a relatively new product, to Fleet (sodium phosphate), a well-studied and widely used preparation. This study was designed to compare the efficacy and patient tolerance of these two bowel preparations for colonoscopy. METHODS: A randomized, single-blinded, prospective trial was conducted. A total of 400 consecutive patients presenting for elective colonoscopy at St George Private Hospital during a 20-week period were randomly assigned to receive Picoprep-3 or Fleet. Patients were asked to record the effects of the preparation, noting tolerability, taste, and side effects. Two hundred patients were assigned to the Picoprep-3 group and 200 to the Fleet group. Surgeons were blinded to the preparation used and rated the quality of the bowel preparation on a scale of 1 to 5 (1 being the optimal score). RESULTS: Picoprep-3 was found to be better tolerated (P < 0.0001) and better tasting (P < 0.0001) than Fleet. Patients in the Picoprep-3 group reported significantly less nausea (P < 0.001), vomiting (P < 0.004), dizziness (P < 0.01), abdominal pains (P = 0.0005), and thirst (P < 0.0001) associated with the preparation. There was no significant difference in visualization of the colon between the two groups as judged by the two colonoscopists (P = 0.06). CONCLUSIONS: Colonoscopy preparation with Picoprep-3 has similar efficacy but superior taste and tolerability compared with Fleet. Picoprep-3 caused less adverse side effects in the study population. PMID- 15043298 TI - Successful low-dose chemotherapy using vinblastine and methotrexate for the treatment of an ileoanal pouch mesenteric desmoid tumor: report of a case. AB - The purpose of this report was to describe the first known case of ileoanal pouch salvage by a low-dose regimen of vinblastine and methotrexate chemotherapy for the treatment of desmoid tumor arising from the mesentery of the ileoanal pouch in a patient who had undergone ileal pouch-anal anastomosis for familial adenomatous polyposis. Mesenteric desmoid tumor involving the ileoanal pouch in a 28-year-old female was treated with vinblastine and methotrexate biweekly for 12 months and monthly for 12 months in an outpatient unit. The desmoid tumor response to the treatment was assessed at routine intervals by physical examination and magnetic resonance imaging. Desmoid tumor was successfully treated with a low-dose regimen of vinblastine and methotrexate chemotherapy without significant side effects, and function of the ileoanal pouch was fully preserved. Magnetic resonance imaging showed a decrease in desmoid tumor size and cellularity, and changes consistent with fibrosis. This is a unique case highlighting the possibility of ileoanal pouch salvage by low-dose combination chemotherapy using vinblastine and methotrexate in a familial adenomatous polyposis patient with mesenteric desmoid tumor. PMID- 15043299 TI - Intrapelvic-perianal hydatid disease in an unusual location: report of a case. PMID- 15043300 TI - Fibrin glue treatment of low rectal and pouch-anal anastomotic sinuses. AB - PURPOSE: This report describes a treatment method for patients with persistent anastomostic sinuses in which fibrin glue is used. METHODS: A retrospective review was conducted of the medical records of seven patients with radiologically documented sinus tracts after restorative proctocolectomy or low rectal anastomosis was managed with fibrin glue obliteration of the tract. The sinus was gently debrided with a curette and then filled with fibrin glue. Postoperatively, the patients received metronidazole 1.5 g per day in divided doses for one week. Outpatient examination of the internal opening was performed at 1, 3, and 12 weeks postoperatively. RESULTS: In all patients, healing of the sinus was observed after one week. After an average of 11.2 months (range, 3-15) of follow up there were no recurrences and no episodes of pelvic sepsis. CONCLUSION: On the basis of this experience, we believe that fibrin glue injection may be an alternative method of managing pelvic anastomotic sinuses. PMID- 15043301 TI - Merkel cell carcinoma of the anal canal: importance of radiotherapy. PMID- 15043302 TI - Botulinum toxin A in anal fissure: why does it work? PMID- 15043303 TI - Clinicopathologic analysis of early-stage sporadic ovarian carcinoma. AB - The reported experience with early-stage (FIGO stage I/II) ovarian carcinoma (OC) is limited given that the majority of women with OC are diagnosed at an advanced stage. There has not been an extensive review of these tumors, and since the pathologic criteria differentiating invasive and borderline tumors have evolved over time, the issue of whether a proportion of these tumors should be reclassified has not been addressed. We identified patients with stage I/II invasive OC who underwent primary surgical management at Memorial Sloan-Kettering Cancer Center from 1980 to 2000. Patients known to have a BRCA mutation or a family history of breast/ovarian cancer were excluded. Hematoxylin and eosin slide review, blinded to clinical outcomes, using current diagnostic criteria for ovarian carcinomas and borderline ovarian tumors, was performed. Progression-free survival (PFS) and disease-specific survival (DSS) were estimated and compared. Hematoxylin and eosin slides were reviewed for 140 of the 145 patients identified. The diagnosis was changed to borderline (low malignant potential) in 41 cases (29.3%). Twenty-nine (70.7%) of 41 changes in diagnosis involved endometrioid and mucinous tumors. This was attributable to the application of recently revised criteria for distinguishing borderline tumors from carcinomas. None of the originally diagnosed clear cell carcinomas was reclassified as borderline. The distribution of histologic subtypes among the 94 carcinomas included 26 serous (27.7%), 25 clear cell (26.6%), 22 endometrioid (23.4%), 10 mixed (10.6%), 6 mucinous (6.4%), 2 malignant Brenner (2.1%), and 3 adenocarcinomas, not otherwise specified (3.2%). Adjuvant therapy was given to 84 (89.4%) of the 94 patients with carcinomas. The 5-year PFS and DSS were significantly greater for the group of cases that was reclassified as borderline (4.5% vs. 26.2% progressed [P = 0.006]; 4.5% vs. 25.6% died [P = 0.003]). The 5 year PFS and DSS were significantly worse for carcinomas with a TP53 mutation (22.6% vs. 41.2% progressed [P = 0.04]; 21.7% vs. 24.7% died [P = 0.04]). There were no statistically significant differences in outcome between stages I versus II, tumor grades, clear cell histology versus other, and stage IC preoperative versus intraoperative rupture. We concluded that a large number of cases originally diagnosed as early-stage sporadic OC were borderline tumors. Clear cell histology does not confer a worse prognosis compared with other histologies. The presence of a TP53 mutation was an adverse prognostic indicator. PMID- 15043304 TI - Distinction of endocervical and endometrial adenocarcinomas: immunohistochemical p16 expression correlated with human papillomavirus (HPV) DNA detection. AB - Determining the origin of uterine adenocarcinomas can be difficult in biopsy and curettage specimens because the morphologic spectrum of endocervical and endometrial adenocarcinomas overlaps. In hysterectomy specimens, the primary site is often equivocal for tumors that involve the lower uterine segment and endocervix and lack identifiable precursor lesions. Most endocervical adenocarcinomas (ECAs) contain high-risk human papillomavirus (HPV) DNA, whereas endometrial adenocarcinomas (EMAs) rarely do. p16 is an inhibitor ofcyclin dependent kinases, and overexpression of p16 has been observed in cervical intraepithelial lesions and invasive carcinomas associated with high-risk HPV types. We evaluated the utility of immunohistochemistry for p16 in the distinction of ECAs and EMAs. p16 expression was assessed in 24 unequivocal EMAs and 19 unequivocal ECAs and correlated with HPV DNA detection by in situ hybridization and polymerase chain reaction. These assays were then used to assist in the classification of four lower uterine segment/upper endocervical adenocarcinomas (LUS/EC-A) of equivocal origin. p16 expression was moderate strong and diffuse in 18 ECAs (median 90% of tumor cells positive, range 90% 100%), and weak and diffuse in one. Fourteen of these were positive for HPV DNA, whereas 5 lacked detectable HPV DNA by in situ hybridization; one of these 5 was positive by polymerase chain reaction. In contrast, EMAs displayed weaker staining with patchy distribution (median 30% of tumor cells positive, range 5% 70%) and none contained HPV DNA by in situ hybridization. Two LUS/EC-As, which were positive for HPV, exhibited strong, diffuse p16 expression, consistent with endocervical origin of the tumors. The remaining 2 LUS/EC-As showed patchy p16 staining and did not contain detectable HPV DNA, consistent with the endometrial origin of the tumors. The p16 expression pattern can distinguish ECAs from EMAs. Compared with HPV DNA detection by in situ hybridization, p16 immunohistochemistry appears to be a more sensitive and easier to perform method for distinguishing ECAs from EMAs, can be used to assist in the classification of LUS/EC-As of equivocal origin, and should be evaluated for its utility in the prospective classification of uterine adenocarcinomas in curettage specimens prior to hysterectomy. PMID- 15043305 TI - CD44 loss in gastric stromal tumors as a prognostic marker. AB - BACKGROUND: The adhesion molecule CD44 (CD44s; CD44H) and its isoforms (CD44v3-6 and v9) are preferentially expressed by different cell types. These transmembrane glycoproteins are involved in cell-cell and cell-matrix interactions and in cell trafficking and, thus, may play a role in tumor metastasis and/or local invasion. The expression pattern of CD44s and variant isoforms, particularly CD44v6 and CD44v9, of some neoplasms, including soft tissue tumors, correlates with clinical course and outcome. The clinical behavior of gastrointestinal stromal tumors (GIST) is site specific; however, other reliable predictors of clinical outcome have not been identified. Thus, the prognostic value of CD44s and isoform expression in GIST were evaluated by immunohistochemistry of tissue microarrays. DESIGN: Paraffin-embedded formalin-fixed tissue cores (129: 103 GIST and 26 normal stomach smooth muscle) from 33 patients with clinical outcome data were collected and used for the construction of the tissue microarrays. One to five tissue cores from each patient specimen were evaluated (mean = 3 tissue cores/patient). Array slides were stained with anti-CD44s (CD44H) and with antibodies to v3, v4, v5, v6, and v9 isomers. CD44s and isoform expression and staining intensity were scored semiquantitatively without knowledge of patient identity or outcome: 0 = no; 1 = weak; 2 = moderate; 3 = moderate to strong; 4 = strong. The scores of multiple cores from the same GIST were averaged; the nonneoplastic smooth muscle was similarly graded. CD44s and isoform expression and intensity were compared with outcome. RESULTS: The 33 patients with gastric GIST, 0.8 to 30 cm in size, were followed for 1 to 111 months with a median follow-up of 7 months (mean 17.5 months). The overall median survival was 25 months. Nine of the 33 (27%) patients had metastases, 9 (27%) had recurrent disease, and 9 (27%) died of disease (9-111 months; mean 39 months; median 23 months). All 18 patients with GIST CD44s expression > 2+ were alive at last follow-up (1-62 months; median 3.5 months; mean 11 months). More than half (53%) of patients with GIST CD44s expression < or = 2+ died (9-111 months; median 23 months; mean 38 months); the median follow-up of the surviving patients with CD44 expression < or = 2 was 5 months (2-22 months; mean 6.5 months; log rank P = 0.07). The majority of tumors were variably positive CD44v3 and CD44v4, but there was minimal staining (number of cases and/or expression level) with antibodies directed against the v5, v6, and v9 isomers. CONCLUSION: These preliminary results suggest that although gastric GISTs variably express CD44s and variants, only the expression of CD44s correlates with clinical outcome with loss of CD44s positivity correlating with poor clinical outcome. PMID- 15043306 TI - Leiomyosarcoma of the kidney: a clinicopathologic study. AB - Leiomyosarcoma of kidney is a rare lesion for which limited data are available. Cases coded as leiomyosarcoma of the kidney from three institutions were reviewed. These cases comprised 3 men and 7 women, 40 to 75 years of age. Immunohistochemistry was performed where possible and clinical follow-up information was obtained. On immunohistochemical staining, 6 of 6 tumors expressed smooth muscle actin, desmin, calponin, and h-caldesmon, and epithelial membrane antigen was positive in 1 of 5. Tumors were negative for cytokeratin and S-100 protein. Follow-up information was available for 9 patients. Two patients had metastases at diagnosis, four developed metastases, and two had recurrent disease. Five of 9 died of disease. Two patients were alive with no evidence of disease after 19 and 60 months, and 2 patients were alive with disease after 48 months and 56 months. Comparing outcome with tumor grade, the one patient with grade 1 tumor was alive with no evidence of disease; of 5 patients with grade 2 tumor, 2 died of disease, 1 was alive with no evidence of disease, 1 was alive with disease, and 1 was alive with extensive disease; all 3 grade 3 patients died of disease. In summary, the majority of renal leiomyosarcomas are intermediate or high grade with correspondingly poor prognosis. PMID- 15043307 TI - Patterns of inflammation in mucosal biopsies of ulcerative colitis: perceived differences in pediatric populations are limited to children younger than 10 years. AB - The histologic criteria used to diagnose ulcerative colitis in colonic mucosal biopsies have been established for many years and include crypt architectural distortion, plasmacellular infiltrates, and neutrophils in the crypt epithelium and lumen. In several recent studies, it has been noted that colonic mucosal biopsies from children presenting with ulcerative colitis show fewer histologic abnormalities at initial presentation, especially less architectural distortion, than do biopsies from adults. In this study, colonic mucosal biopsies taken at the time of presentation of ulcerative colitis in 15 adults and 25 children were examined blindly by two pathologists. All biopsies were taken prior to the initiation of therapy. Twelve children were between 1 and 10 years of age, and 13 children were between the ages of 11 and 17 years. All patients had at least 1 year of follow-up, with clinical and pathologic confirmation of the diagnosis of ulcerative colitis. Five separate histologic features that are characteristic of ulcerative colitis were scored on mucosal biopsies. Children < or = 10 years of age had significantly less crypt branching, plasma cells in the lamina propria, cryptitis, crypt abscesses, and epithelial injury than adults (P values ranging from < 0.0001 to 0.0032). Children between the ages of 11 and 17 years had less cryptitis, crypt abscesses, and epithelial injury than adults (P values ranging from 0.0001 to 0.007) but similar degrees of crypt architectural distortion and plasma cell infiltrates. For all histologic features examined except epithelial injury, the significant findings were due to differences in biopsies taken proximal to the rectum. No significant differences in histology scores were found in rectal biopsies between any age group, except for epithelial injury, which was significantly less in children < or = 10 years. The findings show for the first time that the perceived differences between adults and children with ulcerative colitis are largely due to a decrease in histologic features of colitis in children less than 10 years of age. As children approach adulthood, the degree of inflammation and architectural distortion seen is similar to that found in adults. However, rectal biopsies show similar degrees of colitis in all age groups. PMID- 15043308 TI - Pediatric patients with untreated ulcerative colitis may present initially with unusual morphologic findings. AB - BACKGROUND: Anecdotal observations by the authors of this study, together with the results of a few previous smaller studies, suggest that children with new onset (previously untreated) ulcerative colitis (UC) may occasionally present with discontinuous disease, relative or absolute rectal sparing, and may lack histologic features ofchronicity. Therefore, the objectives of this study were to determine the clinical and pathologic features of new-onset UC in children and to compare the initial presentation of UC in this group with a control group of adults. DESIGN: Routinely processed rectal and colonic mucosal biopsies from 73 pediatric (male/female ratio 33/40, mean age 11.5 years, range 2.5-18 years) and 38 adult patients (male/female ratio 15/23, mean age 41.5 years, range 27-64 years) who presented with new-onset UC were evaluated for a variety of clinical and pathologic features, including duration of symptoms prior to presentation, crypt architectural (e.g., atrophy, branching) and nonarchitectural (e.g., basal plasmacytosis, Paneth cell metaplasia) features of chronicity, degree of active inflammation, and distribution and extent of disease. RESULTS: A significant proportion of children with new-onset UC had patchiness of microscopic features of chronicity (21% of patients), relative (23%), or absolute (3%), rectal sparing, and had little or no crypt architectural distortion in their rectal biopsies (8%). These features were not observed in adult patients with UC. In addition, a higher proportion of children with UC initially presented with subtotal or with pancolitis compared with the adults (42% vs. 11%; P < 0.002). CONCLUSIONS: A significant proportion of children with new-onset UC may show unusual patterns of disease. Pathologists should be aware of these findings since they have significant implications for the differential diagnosis of pediatric inflammatory bowel disease. PMID- 15043309 TI - Grade of stromal invasion in small adenocarcinoma of the lung: histopathological minimal invasion and prognosis. AB - The pathologic features of invasion such as stromal disruption and pleural/vascular involvement have been shown to be of prognostic value in adenocarcinoma. However, the relationship between the degree of invasion, histologic subtype of adenocarcinoma, and prognosis remains unclear. We retrospectively studied 380 peripheral adenocarcinomas of < or = 2.0 cm in diameter with regard to histology and clinical profiles. Their degree of invasive growth was classified into four grades as follows according to the structural deformity and its location in the adenocarcinoma lesion: Grade 0 had a pure bronchioloalveolar growth pattern and no evidence of stromal invasion. Grade 1 had stromal invasion in the area of bronchioloalveolar growth. Grade 2 had stromal invasion localized on the periphery of a fibrotic focus. Grade 3 had stromal invasion into the center of a fibrotic focus. The clinicopathological data were obtained from medical records. The distribution of the histologic grade of invasion was as follows: grade 0 in 85 tumors (22%), grade 1 in 37 (10%), grade 2 in 46 (12%), and grade 3 in 212 (56%). This histologic grade of invasion was closely related to other indicators of tumor spread. Vascular/lymphatic permeation was seen in none of grade 0, in 1 lesion each of grade 1 and grade 2, and 144 (68%) of grade 3. Lymph node metastasis was seen in 57 (27%) lesions of grade 3 but not in grades 0, 1, or 2. The 5-year disease-free survival rates were 100%, 100%, 100%, and 59.6% for tumors with grade 0, grade 1, grade 2, and grade 3 invasion, respectively. Tumors with grade 1 and grade 2 invasion, like tumors with grade 0 invasion (bronchioloalveolar carcinoma), showed an excellent prognosis. Therefore, tumors with grade 1 and grade 2 invasion could be considered "minimally invasive" or "early" adenocarcinomas. PMID- 15043310 TI - Pulmonary meningothelial-like nodules: a genotypic comparison with meningiomas. AB - BACKGROUND: Minute pulmonary meningothelial-like nodules (MPMNs) are incidental interstitial pulmonary nodules. They share histologic, ultrastructural, and immunohistochemical features with meningiomas (MGs). DESIGN: Sixteen cases yielding 33 separate MPMNs and 10 cases of benign MG were studied. Immunohistochemical studies and mutational analyses were performed on microdissected tissue using 20 polymorphic microsatellite markers targeting 11 genomic regions in an effort to identify genetic similarities of MPMN and MG. RESULTS: A total of 96.6% of MPMNs stained positive for vimentin, 33.3% for epithelial membrane antigen, 3% for S-100, and all were negative for cytokeratin and synaptophysin. Loss of heterozygosity (LOH) was identified in 25% of single MPMN affecting 3 genomic loci. No solitary MPMN had more than 1 LOH event. Multiple LOHs were seen only in MPMN-omatosis syndrome, where 33.3% of MPMNs showed LOH affecting 7 genomic loci. MG showed the highest frequency of LOH with major events seen at 22q (60%), 14q (42.8%), and 1p (44.4%) that were not shared by MPMN. CONCLUSION: Isolated MPMN lacks mutational damage, consistent with a reactive origin. MPMN-omatosis syndrome might represent the transition between a reactive and neoplastic proliferation. MPMNs are different from MG based on the major molecular genetic events seen in their formation and progression. PMID- 15043311 TI - Patterns of morphologic alteration in residual rectal carcinoma following preoperative chemoradiation and their association with long-term outcome. AB - Preoperative radiation (RT) and chemotherapy improve outcome in patients with locally advanced rectal adenocarcinoma and, therefore, have been used increasingly in patient management. The histopathologic alterations in postirradiated rectal adenocarcinoma and their prognostic significance have not been fully characterized. In this study, detailed analyses of morphologic alterations of stromal and tumor cells were performed in a series of 66 posttreatment rectal carcinomas, and the pathologic findings were correlated with long-term outcome. All tumors were locally advanced, with a bulky and/or tethered tumor or endorectal ultrasound or magnetic resonance imaging evidence of T3-4 and / or N1 disease. All patients were treated at one institution with preoperative RT to the pelvis (at least 4500 cGy) with or without concurrent 5-fluorouracil (5 FU)-based chemotherapy 4 to 7 weeks prior to surgical resection. Pathologic assessment showed some treatment response in all patients. Nine patients (13.4%) had complete response, and 8 (11.9%) had near-complete response (> 95% of the tumor replaced by fibroinflammatory tissue). Salient morphologic features included marked fibrosis with or without prominent inflammatory cells replacing neoplastic glands; lack of active tumor necrosis; increased mucin production and mucin pools; marked cytoplasmic eosinophilia, often in combination with marked nuclear atypia but without active mitoses in tumor cells showing treatment effect; endocrine tumor phenotype; and retention of mucosal adenoma in the presence of tumor regression within the bowel wall. With a median follow-up of 69 months, the estimated 5-year recurrence-free survival (RFS) for the entire group was 79%. By univariate analysis, the residual tumor stage (P < 0.05) and reduction of pretreatment T stage (P = 0.002) significantly correlated with RFS, as did pN stage (P = 0.002) and lymphovascular invasion (P = 0.008). The extent of treatment response did not correlate with RFS (P = 0.4). However, patients with a treatment response > or = 95% seemed to fare better than those with a treatment response < 95% (marginally significant difference in RFS, P = 0.057). Univariate and multivariate analyses identified the following morphologic patterns that were significantly associated with a reduced RFS independent of other risk factors: a fibrotic-type stromal response with minimal inflammatory infiltrates (P = 0.001) and absence of surface ulceration (P = 0.026). Our study represents the first detailed morphologic assessment of rectal carcinomas that have been subjected to long course preoperative RT and chemotherapy. Our results demonstrate distinct morphologic features in treated rectal carcinomas that are prognostically relevant. PMID- 15043312 TI - Molecular detection of JAZF1-JJAZ1 gene fusion in endometrial stromal neoplasms with classic and variant histology: evidence for genetic heterogeneity. AB - Endometrial stromal tumors (ESTs), including low-grade endometrial stromal sarcomas (LGESSs) and endometrial stromal nodules (ESNs) of classic histology, exhibit characteristic morphologic features and contain the nonrandom t(7;17)(p15; q21), which results in the fusion of two novel genes, JAZF1 and JJAZ1. ESTs may pose diagnostic challenges when they involve extrauterine sites, present as metastases, or display variant histologic appearances. The aim of this study was to evaluate the frequency of the JAZF1-JJAZ1 gene fusion among primary uterine, metastatic, and primary extrauterine ESTs of various histologic types and its role as a possible diagnostic adjunct. Using a nonnested reverse transcriptase-polymerase chain reaction approach, we assayed for JAZF1-JJAZ1 gene fusion transcripts in 10 cases with available fresh-frozen tissue. These included five primary uterine (two classic, one mixed smooth muscle, and one epithelioid LGESS; one classic ESN), four metastatic (two fibromyxoid, one classic, and one epithelioid LGESS), and one extrauterine (classic LGESS) tumor. The same primer set and assay conditions were used on five additional paraffin-embedded cases with adequate RNA, including three primary uterine (one fibromyxoid and one mixed smooth muscle LGESS; 1 mixed smooth muscle ESN) and two intraabdominal recurrent (two mixed smooth muscle LGESSs) ESTs. Two cellular leiomyomas and one ESS cell line (ESS-1) without the t(7;17) at the cytogenetic level were run in parallel as controls. JAZF1-JJAZ1 gene fusion transcripts were detected in five (33%) of 15 ESTs, including three of eight primary uterine, one of four metastatic, one of one extrauterine, and none of two recurrent cases. Most ESTs of classic histology showed evidence of JAZF1-JJAZ1 fusion (4 of 5 cases), whereas only one mixed smooth muscle ESN of 10 variant cases was positive. Positivity for JAZF1-JJAZ1 fusion transcripts was found in four of 10 fresh-frozen samples and in one of five paraffin-embedded ESTs. The control specimens were all negative. In conclusion, our data suggest that ESTs are genetically heterogeneous, with the prevalence of the JAZF1-JJAZ1 fusion being highest among ESTs of classic histology. Hence, the diagnostic utility of a JAZF1-JJAZ1 fusion transcript assay in ESTs may be limited to the classic histologic subset. PMID- 15043313 TI - Intraductal tubular adenoma, pyloric type, of the pancreas: additional observations on a new type of pancreatic neoplasm. AB - Three cases of a distinctive intraductal tubular adenoma, pyloric type, of the main pancreatic duct are reported. The patients, two women and a man, whose ages ranged from 63 to 70 years, complained of abdominal pain attributed to chronic pancreatitis in two patients. The patient with the largest tumor also had symptoms of gastric outlet obstruction. The tumors, two of which arose in the head and one in the tail of the pancreas, led to occlusion and cystic dilatation of the main pancreatic duct. Two adenomas were sessile and one was attached to the wall of the pancreatic duct by a thin fibrous stalk. Microscopically, they were composed of lobules of closely packed tubular glands similar to pyloric glands. In one tumor, nearly all glands were lined by columnar mucin-secreting cells with abundant clear cytoplasm and basally oriented nuclei. In addition to pyloric glands, two adenomas contained glands lined by cells with little or no mucin as well as by pink oncocytic cells. Focal intestinal differentiation was identified in one tumor. Both intracellular and extracellular mucin was detected with the mucicarmine, periodic acid-Schiff, and alcian blue stains. All three adenomas were CK7 positive and CK20 negative. Focal carcinoembryonic antigen linear reactivity along the apical cytoplasm was seen in many cells, but few cells expressed cytoplasmic carcinoembryonic antigen. All three adenomas showed low proliferative activity as measured by the MIB-1 labeling index. The three adenomas were p53 negative and showed loss of DPC4 expression. No endocrine cells were identified in any of the tumors. All patients are alive and symptom free from 4 months to 5 years following surgical treatment. PMID- 15043314 TI - How often does alpha-methylacyl-CoA-racemase contribute to resolving an atypical diagnosis on prostate needle biopsy beyond that provided by basal cell markers? AB - BACKGROUND: Alpha-methylacyl-CoA-racemase (AMACR), a recently discovered tumor marker for prostate cancer, is being used increasingly in conjunction with hematoxylin and eosin (H&E) histology and basal cell markers in the workup of difficult prostate needle biopsies. However, it is not known how often a positive AMACR staining is used merely to support a malignant diagnosis that could otherwise be established based on routine H&E histology and negative basal cell staining. METHODS: This study included 307 prostate needle biopsies that were sent to us for consultation diagnosed as "atypical" by contributing pathologists. Immunohistochemistry for AMACR, high molecular weight cytokeratin, and p63 was performed. AMACR staining intensity was graded as negative, weak, moderate, and strong. Only staining that was significantly stronger than that of background benign glands was considered positive. An "expert review" diagnosis was first rendered as benign, atypical, or cancer based on the H&E-stained section and basal cell marker stains without the knowledge of AMACR expression. The AMACR stains were then reviewed and a final diagnosis was rendered. RESULTS: A total of 215 cases had a final diagnosis of cancer following evaluation of the H&E-stained section, basal cell markers, and AMACR. Of these 215 cases, 176 (81.9%) were positive and 39 (18.1%) were negative for AMACR staining. Of 81 cases with a final diagnosis of atypical following review of all material, 42 (51.9%) were positive and 39 (48.1%) were negative for AMACR staining. When AMACR staining was negative, in no case was the initial cancer, atypical, or benign diagnosis (based on routine histology and negative basal cell markers) changed based on AMACR stain results. Of 115 cases called atypical after expert review, 76 were positive for AMACR; of these 76 cases, 34 (44.7%) were changed to a final diagnosis of cancer. AMACR expression was positive, with moderate and strong staining in 30 of the 34 cases (88.2%), for which the immunohistochemical result converted the expert review atypical diagnosis to a final cancer diagnosis. Of these 34 cases, 11 underwent radical prostatectomy, and cancer was found in all cases. Three additional patients underwent repeat biopsy, and cancer was present in the repeat biopsy in 2 patients. The cases whose diagnosis was changed from "atypical" on expert review to cancer were all highly suspicious for cancer based on H&E histology and negative basal cell markers, yet a definitive cancer diagnosis could not be established because of small size, insufficient cytologic atypia, or biopsy artifact. CONCLUSION: Interpretation and use of AMACR staining should be executed with caution. A negative AMACR stain can be seen in approximately 18% of cases considered to be cancer based on H&E stain combined with negative basal cell markers. A positive AMACR staining converted an atypical diagnosis, based on suspicious histology and negative basal cell marker stains, to cancer in approximately 10% (34 of 307) of cases thought to be atypical by contributing pathologists and in approximately 50% (34 of 76) of cases thought be atypical on expert review by a specialist in genitourinary pathology. PMID- 15043315 TI - Uterine epithelioid leiomyosarcomas with clear cells: reactivity with HMB-45 and the concept of PEComa. AB - In this study, we investigated HMB-45 expression in epithelioid uterine leiomyosarcomas with clear cell areas. From 12 epithelioid leiomyosarcomas, we selected 5 that had: 1) clear cell areas and 2) spindle cell areas that were at least focally positive for desmin and caldesmon. The patients' ages ranged from 47 to 82 years (mean 64 years). Presenting symptoms were uterine bleeding (three), abdominal pain (one), and a pelvic mass (one). There was no history of tuberous sclerosis or lymphangioleiomyomatosis. One patient had stage II disease, one stage III, and three stage IV. All were treated with total abdominal hysterectomy and bilateral salpingo-oophorectomy. Two received radiotherapy, and three were also treated with chemotherapy. The tumors ranged in size from 4 x 3 x 3 cm to 10 x 7 x 6 cm; all had significant cellular atypia, areas of coagulative necrosis, and between 10 and 90 mitoses per 10 high power fields. Vascular invasion was seen in three cases. The epithelioid component varied from 50% to 90% in each case; and the percentage of clear cells was < 1% in one case, 5% in one case, and 10% to 80% in three cases. Smooth muscle actin and desmin were positive in all cases. Four cases were positive for HMB-45 only in the clear cell areas. The tumor with < 1% of clear cells was negative for HMB-45. All were negative for S-100 and c-kit. Three patients died of disease at 9, 30, and 32 months; one patient is alive with progressive disease at 6 months, and one patient (stage II disease) is alive with no evidence of disease at 8 months. Unequivocal uterine epithelioid leiomyosarcomas may have clear cells positive for HMB-45. These tumors might belong to the group of lesion designated as PEComas; however, it is advisable to designate them as uterine leiomyosarcomas. In uterine smooth muscle tumors, some epithelioid cells most likely undergo clear cell changes and become positive for HMB-45. It would be advisable to perform this stain in all epithelioid smooth muscle tumors of the uterus. PMID- 15043316 TI - Hepatoid thymic carcinoma: report of a case. AB - We describe the clinicopathologic findings in a so far unrecognized thymic tumor. The tumor occurred in a 70-year-old woman with respiratory distress but neither myasthenia gravis nor other symptoms. Metastases or another primary tumor were absent. The well-circumscribed neoplasm was located in the thymic region, measured 18 x 12 x 8 cm, and showed a homogeneous, tan-colored, soft cut surface. By histology, the tumor lacked a true capsule and a lobular growth pattern, was almost devoid of stroma, and infiltrated among remnant thymus lobules. The polygonal tumor cells formed solid sheets, trabeculae, or occurred as single cells that resembled hepatocytes. Proliferative activity was low. Portal structures, sinuses, and bile were absent as were areas of conventional thymoma, adenocarcinoma, or germ cell tumor. The tumor expressed cytokeratins 7 and 19, alpha1-antitrypsin, alpha1-antichymotrypsin, and hep-Par-1. Alpha-fetoprotein (AFP), human beta-chorionic gonadotropin (beta-HCG), placental alkaline phosphatase, CD5, CD30, CD31, CD34, CD45, CD68, CD99, S-100, HMB45, desmin, actin, or neuroendocrine markers were not expressed, and intratumorous CD1a+ or TdT+ immature T cells were absent. AFP was repeatedly undetectable in the blood. Mediastinal tumor recurrence was detected 6 months after surgery. Following radiochemotherapy, the patient has remained free of disease for 26 months. We conclude that this tumor is a thymic carcinoma (WHO type C thymoma). A diagnosis of hepatoid yolk sack tumor appears unlikely considering absence of a bona fide germ cell component, lack of AFP expression, and the patient's female gender. Because of its morphologic and immunohistochemical features, we propose the term "hepatoid thymic carcinoma" for this new type of thymic carcinoma. PMID- 15043317 TI - Reactive angioendotheliomatosis of the intestine. AB - We present a case of reactive angioendotheliomatosis (RAE) of the colon, featuring intravascular proliferation of endothelial cells with histologic resemblance to glomeruloid hemangioma. A 19-year-old Japanese male with an anal fistula was diagnosed endoscopically with Crohn's disease. Six months later, he was hospitalized for fever and abdominal pain. Emergency resection of ileocecum and splenic flexure of the colon was undertaken to control massive intestinal hemorrhage, and in all parts of the resected colon, foci of many small vessels with intravascular proliferation of endothelial cells were noted throughout the layers. Moreover, solid proliferation of endothelial cells was seen in the submucosa at the base of open ulcers. Two small granulomas, compatible with Crohn's disease, were also evident in the muscle layer of the terminal ileum. No other hemangiomas or hemangioma-like structures were observed with CT scans, and the vascular lesions were histologically diagnosed as RAE. The pathogenesis of this disorder is unknown, and most cases occur in skin with systemic disease. The present case might thus be a first case of RAE of the intestine without cutaneous involvement. Whether there is a relation with coexistent enteritis suggestive of Crohn's disease needs to be clarified. PMID- 15043318 TI - Severe malabsorption due to refractory celiac disease complicated by extensive gastric heterotopia of the jejunum. AB - Refractory celiac disease denotes that patients considered to have celiac disease fail to respond histologically to treatment with a gluten-free diet. Among several causes of nonresponsiveness, enteropathy-type T-cell lymphoma is most important because of its almost invariably rapid lethal outcome. We present the case of a 44-year-old patient with refractory celiac disease complicated by unusually severe malabsorption. Repeated duodenal biopsies disclosed normal and slightly shortened villi, focal crypt hypertrophy, and a moderate increase of intraepithelial lymphocytes consistent with celiac disease, but unable to explain the severe malabsorption. To rule out cryptic lymphoma, push enteroscopy was done providing 21 biopsies taken along the entire jejunum. Surprisingly, about 70% of the biopsies were composed of gastric glands covered by nonabsorptive-type, strongly periodic acid-Schiff-positive surface epithelium and showed a villous architecture. Alternating with the gastric mucosa, there were areas of flat mucosa with elongated crypts and occasional erosions. Irrespective of the type of surface epithelium, intraepithelial lymphocytes were increased with counts up to 80/100 epithelial cells. Despite harboring an aberrant immunophenotype, overt T cell lymphoma was ruled out histologically and by lack of monoclonality, as tested by polymerase chain reaction. To the best of our knowledge, this is the first case of refractory celiac disease complicated by extensive jejunal gastric heterotopia, which might have contributed to the severe malabsorption. PMID- 15043319 TI - Intraductal carcinoma of the oral cavity: a case report and a reappraisal of the concept of pure ductal carcinoma in situ in salivary duct carcinoma. AB - The pure in situ form of salivary duct carcinoma, also known as intraductal carcinoma, is very rare, and its existence is controversial. We describe a case arising from the minor salivary glands. The patient was a 44-year-old woman who presented with a painless mass in the buccal mucosa. On microscopic examination, the tumor comprised crowded and smooth-contoured epithelial units exhibiting a fenestrated or cribriform pattern, occasionally punctuated by comedonecrosis. An attenuated layer of myoepithelial cells could be demonstrated around all the islands by immunostaining for p63 and actin, indicating absence of an invasive component. The patient remained well following local excision. This case, together with other reported cases, suggests that intraductal carcinoma is a distinct entity. It may represent the preinvasive phase of some invasive salivary duct carcinomas but by itself is nonmetastasizing and associated with an excellent prognosis. PMID- 15043321 TI - International variation in histologic grading is large and persistent feedback does not improve reproducibility. PMID- 15043320 TI - Whorls of intermediate filaments with entrapped neurosecretory granules correspond to the "rhabdoid" inclusions seen in pancreatic endocrine neoplasms. PMID- 15043322 TI - Histologic localization of sentinel lymph node metastases in breast cancer. PMID- 15043323 TI - Reactive nodular fibrous pseudotumor of the gastrointestinal tract and mesentery. PMID- 15043324 TI - Multiple small intestinal carcinoid tumors: independent primary neoplasms or metastases? PMID- 15043325 TI - Detecting and preventing colorectal cancer in specific communities. PMID- 15043326 TI - Patterns of colorectal cancer incidence, risk factors, and screening in Kentucky. AB - BACKGROUND: Colorectal cancer incidence rates are higher in Kentucky than in the United States in general, and there are regional variations within the state. METHODS: This study investigates these variations in relation to lifestyle and health behaviors, combining data from the Kentucky Behavioral Risk Factor Surveillance System (BRFSS), and from the Kentucky Cancer Registry. We used Kentucky's fifteen Area Development Districts (ADDs) as units of analysis across a five-year period from 1993 to 97. RESULTS: Differences were observed across ADDs. ADDs with a higher prevalence of risk factors, with the exception of chronic alcohol drinking, had lower CRC rates. ADDs with a higher proportion of respondents having had recent routine check-ups had higher CRC incidence rates. CONCLUSIONS: In general, healthier lifestyles and positive health-related behaviors were associated with increased colorectal cancer incidence. This may be explained by the tendency for healthier individuals to receive regular check-ups and screening, thus increasing the detection rate of colorectal cancer. PMID- 15043327 TI - Screening preferences for colorectal cancer: a patient demographic study. AB - BACKGROUND: Colorectal cancer is a leading cause of cancer-related death. Screening for colorectal cancer is a rational and cost-effective strategy for reducing the incidence of colorectal cancer and related mortality. Despite endorsement by academic and health care organizations, patient awareness and compliance with screening is low, partly because of patient-related barriers to screening. METHODS: A convenience sample of adults attending the internal medicine and family practice clinics of a community teaching hospital was studied. A description of fecal occult blood testing (FOBT), sigmoidoscopy, and colonoscopy procedures was given in a packet along with a questionnaire. The questionnaire focused on screening procedures followed in our hospital (i.e., yearly FOBT and sigmoidoscopy every 5 years or colonoscopy every 10 years for average-risk individuals). RESULTS: Of the 193 patients who responded, 55% preferred sigmoidoscopy and FOBT, 29% chose colonoscopy, and 16% wanted no screening. Those with knowledge of someone with colon cancer or colon polyps reported a significantly higher preference for screening than those without such knowledge. Catholics were most likely to prefer no screening compared with non Catholics. Ex-smokers (compared with all others) were more likely to want screening. Catholics were least likely to want colonoscopy. Patients with previous experience of colorectal screening preferred future screening. Those preferring no screening were significantly younger than those who expressed a preference for screening. CONCLUSION: The results of this study demonstrate diversity in patient choices for colorectal cancer screening. A focus on people's preferences rather than on the test itself may help develop and target appropriate intervention for prevention of colorectal cancer. PMID- 15043328 TI - Colorectal carcinoma in young females. AB - BACKGROUND: We sought to study the clinicopathologic characteristics of colorectal cancer in young female patients. We also wanted to determine the association of colorectal cancer with anemia in these female patients and, finally, to determine the effect of gender on prognosis in young patients with colorectal cancer. METHODS: We performed a retrospective analysis of all young patients diagnosed with colorectal cancer between 1982 and 1999 in two teaching hospitals in New York City. RESULTS: A total of 3,546 cases of colorectal cancer were diagnosed. Sixty-one (1.63%) of these patients were young patients and 32 (0.85%) were female. Young refers to all patients in the study who were younger than 40 years of age. The clinical presentation and mean age at presentation were very similar in both male and female patients. At presentation, 87.5% of female patients had anemia compared with only 69% of male patients. Males had a statistically significant higher mean hemoglobin level compared with females (12.87 versus 10.29 g) at P = 0.0001. Seventy-nine percent of female patients compared with 86% of male patients presented with left-sided tumors. Fifty-five percent of males presented with late stage disease compared with 68% of females (P = 0.27). Female sex seemed to adversely affect the prognosis, although this did not reach statistical significance (P = 0.08). Stage of disease was associated with worse prognosis and this was independent of sex. Age and hemoglobin were not independent predictors of mortality. CONCLUSION: Colorectal cancer does occur in females of childbearing age who might have a tendency to present with late stage disease as evidence from this study. Young female patients with anemia should be questioned about gastrointestinal symptoms, and colorectal cancer should definitely be in the differential diagnoses. This might conceivably allow for earlier diagnosis and potential for cure in this patient group. PMID- 15043329 TI - Absolute CD4 and CD8 counts and CD4-to-CD8 ratios in eight patients with indolent B-cell chronic lymphocytic leukemia. AB - BACKGROUND: There are some patients with B-cell chronic lymphocytic leukemia who exhibit an extraordinary natural resistance to this malignancy, which lasts for many years. In this study, we report the T-cell subset values and ratios in eight such patients. METHODS: Impath (New York, NY) evaluated immunophenotyping by performing flow cytometry. Absolute CD4 and CD8 counts and CD4:CD8 ratios were performed at Memphis Pathology Laboratory, Memphis, Tennessee. RESULTS: CD4 and CD8 counts and CD4:CD8 ratios were normal in all eight patients, in contrast to the suppressor cell proliferation and low helper-suppressor ratios that have been previously reported in other patients with B-cell chronic lymphocytic leukemia. CONCLUSION: These results require further study to determine their significance. Implications for further study are discussed. PMID- 15043330 TI - Chronic lymphocytic leukemia in Turkey: experience of a single center in Istanbul. AB - BACKGROUND: In this study, the clinical characteristics, survival, and prognostic factors of 200 patients diagnosed as having chronic lymphocytic leukemia (CLL) were analyzed. METHODS: The medical charts of 200 CLL patients registered to our center between 1984 and 2000 were retrospectively evaluated. RESULTS: Of all patients, 129 were men and 71 were women (male/female ratio, 1.82). The median age at the time of initial diagnosis was 63 years (range, 38-90 years). Sixty patients were classified as Binet's Stage A, 49 as Stage B, and 91 as Stage C. Sixty-two cases were diagnosed during routine laboratory examinations when they were asymptomatic. Forty-three patients were lost to follow-up, and 157 patients have been followed regularly until the end of the study period. Hemolytic anemia developed in nine (5.7%) patients, second primary cancer in six (3.8%), and Richter's syndrome in two (1.2%). Forty-eight percent of CLL patients were treated immediately after initial diagnosis. The overall response (complete or partial) to first-line and second-line therapies was 61.6% and 54.4%, respectively. The median time of follow-up for patients followed up regularly was 47 months (range, 1-195 months). Sixty-three patients died during the follow-up: the deaths of 39 (62%) of these were attributable to CLL-related causes. The median survival time was 48 months. The 5-year survival rate was 36.5% and the 10 year survival rate was 8%. Stage according to Rai's classification, lymphocyte count, and age showed a significant prognostic effect on survival by univariate analysis. On multivariate analysis, advanced age and lymphocyte count were independent prognostic parameters. CONCLUSION: In our study, more asymptomatic CLL patients have been diagnosed in recent years. The survival, especially of our early-stage patients, was shorter than that in other CLL series of Western origin. Rai's staging system was seen to determine prognosis better than Binet's staging system. PMID- 15043331 TI - Metastatic spine tumors. AB - Cancer is the second leading cause of death in the United States, and vertebral body metastases often occur in systemic malignancy. Metastatic spinal tumors may present with pain or neurologic deficit, or may be detected during screening examinations in patients with known malignancy. Management of spinal metastases remains controversial. The role of surgery, especially decompressive laminectomy without stabilization, has been questioned. Recent series attest to the beneficial role of surgery, emphasizing anterior and combined decompression and stabilization procedures. We review the relevant literature on metastatic spinal tumors, assessing imaging strategies, adjuvant treatment, patient selection, and results and complications. Operative decompression and stabilization is an important tool in the management of spinal metastatic disease. Patient selection and appropriate use of anterior and/or posterior decompression and stabilization are necessary to optimize surgical results. PMID- 15043332 TI - Management strategies for patients with brain metastases: has radiosurgery made a difference? AB - BACKGROUND: Brain metastases develop in up to 50% of all patients with cancer. The purpose of this retrospective study was to compare the outcomes for brain metastases patients treated with adjuvant whole-brain radiotherapy (WBRT) or stereotactic radiosurgery (SRS). METHODS: Between 1990 and 1995, 86 patients with brain metastases received external beam WBRT with a megavoltage beam at our center. Between January 2000 and July 2001, 48 patients with 84 tumors diagnosed as cerebral metastases were treated with SRS using the Leksell Gamma Knife. A comparative analysis of the outcomes in the two different groups was made to determine a possible statistically significant difference in survival. RESULTS: In the WBRT group, the median follow-up was 11 months. Thirty-nine patients (45.3%) experienced an improvement in neurologic status. The median overall survival was 5 months (range, 1-40 months). In the SRS group, the median follow up for this group was 9 months. Thirty-four patients experienced an improvement in their neurologic signs and symptoms. The median survival was 12 months (range, 1-16 months). CONCLUSION: SRS appears to be a safe and effective treatment option for those patients with a limited number of brain metastases and in patients with controlled or limited systemic disease. PMID- 15043333 TI - Breast cancer incidence and exposure to environmental chemicals in 82 counties in Mississippi. AB - OBJECTIVE: Breast cancer is a major public health problem in the United States and in most industrialized countries. Environmental exposure to several chemicals has been implicated in the cause of breast cancer. However, data are not consistent about the role of the environment in breast cancer incidence. To assess environmental risks for breast cancer, patterns of breast cancer incidence in relation to environmental chemicals in Mississippi counties were mapped. METHODS: This article presents an analysis of age-adjusted incidence rates of female breast cancer and environmental chemical emissions by county in the state of Mississippi. The incidence data were obtained from the State Department of Health, and emissions data sources included the U.S. Environmental Protection Agency's Air-Data and Toxics Release Inventory Program. RESULTS: Counties having a significantly higher incidence of breast cancer compared with the state's rate were identified. Also identified were counties with higher levels of chemical emissions. The incidence of breast cancer in 1998 was significantly associated with the amount of ammonia (r = 0.268, P = 0.015), minimum emissions (r = 0.233, P = 0.035), and maximum emissions (r = 0.237, P = 0.032) of the facilities in the county. A linear dose-response relationship was observed between increased amounts of chemical emissions and breast cancer incidence. CONCLUSIONS: This study found a significant link between breast cancer incidence and maximum emissions of environmental chemicals. However, further research using individual level data is needed. PMID- 15043334 TI - Breast and cervical cancer mortality in the Mississippi Delta, 1979-1998. AB - BACKGROUND: Historically, the Mississippi Delta region has been medically underserved. This may lead to lower cancer prevention efforts and higher breast or cervical cancer mortality rates. METHODS: Death rates for 1979 through 1998 were calculated for Mississippi Delta women and for women living elsewhere in the US. RESULTS: Breast cancer mortality in the Delta was similar to that elsewhere in the US in recent years for both black and white women, but rates were lower in the Delta in the early years of the study period. Overall, cervical cancer mortality was similar in the two areas but rates declined more rapidly elsewhere in the US than in the Delta. Breast and cervical cancer mortality was higher among black women than among white women in both areas. Cervical cancer mortality was higher among white rural and black urban women in the Delta than their counterparts elsewhere. CONCLUSION: These results can guide prevention activities for reducing mortality from these diseases. PMID- 15043335 TI - Association between Epstein-Barr virus and classic Hodgkin lymphoma in Jordan: a comparative study with Epstein-Barr virus-associated Hodgkin lymphoma in North America. AB - An association between Epstein-Barr virus and Hodgkin lymphoma has been shown in several parts of the world. The reported incidence of Epstein-Barr virus in Hodgkin lymphoma varies significantly from one country to another and ranges from <30% in Swedish patients to 100% in patients from Kenya. Using in situ hybridization for detection of Epstein-Barr virus-encoded RNA and immunohistochemistry for detection of Epstein-Barr virus latent membrane protein, we analyzed 28 cases of Hodgkin lymphoma from Jordan and 30 cases from the United States. Eight of 28 Jordanian cases and 9 of 30 North American cases were Epstein Barr virus positive. Our studies show that the incidence of Epstein-Barr virus among Jordanian patients with Hodgkin lymphoma is similar to the rate in patients from the United States. This rate appears to be low to intermediate compared with rates in other parts of the world. PMID- 15043336 TI - Epidural intracranial abscess as a complication of frontal sinusitis: case report and review of the literature. AB - Two cases of epidural abscess as a complication of frontal sinusitis are presented. The diagnoses were suspected on the basis of history and were confirmed by magnetic resonance imaging and computed tomography. Both patients were treated successfully by means of surgery and intravenous antibiotics. One patient developed meningitis in the postoperative course and was treated by changing the antibiotic regimen. However, further follow-up in the outpatient clinic by physical examinations and brain computed tomography scans showed no longterm neurologic complications in either case. Intracranial suppuration, including epidural abscesses, can complicate acute and chronic frontal sinusitis. These complications are diagnosed by maintaining a high index of suspicion and using the appropriate neuroimaging studies without delay. PMID- 15043337 TI - Utilization review and managed health care liability. AB - This article explores the development of jurisprudence interpreting application of the Employee Retirement Income Security Act of 1974 to patient care denials by managed care. It identifies quality-of-care protections for patient care under present federal law. If an insurance company utilization review denies care based on patient-specific reasoning, then the patient may have recourse against the utilization review on the basis of a state law claim of malpractice grounded in medical decision-making by the insurance company. PMID- 15043338 TI - Jaundice secondary to isolated porta hepatis metastasis in colorectal cancer: case report and review of the literature. AB - Colorectal cancer occurs mainly after the age of 50. The liver is the most frequent site of metastases, although isolated metastases to the porta hepatis are rarely reported in the literature. From 1924 to 1993, only 16 cases of periportal lymph nodes metastases were reported. We report a case of jaundice secondary to porta hepatis metastases from primary colorectal cancer. The appearance of symptoms was concurrent with the elevation of carcinoembryonic antigen in our case. This emphasizes the importance of polymerase chain reaction to detect the small amount of carcinoembryonic antigen transcript in blood or in peritoneal fluid before the appearance of symptoms. Polymerase chain reaction allows the prediction of high risk of recurrence and the presence of micrometastases. More trials are needed to assess the outcome after treatment by adjuvant chemotherapy for micrometastases. PMID- 15043339 TI - Chylothorax as a result of chronic lymphocytic leukemia: case report and review of the literature. AB - Chylothorax occurs when a disruption in the thoracic duct allows chyle to escape into the pleural space. The most commonly reported cause is malignancy, especially lymphoma. However, chylothorax caused by chronic lymphocytic leukemia is rarely reported in the literature. We describe a patient who developed chylothorax secondary to chronic lymphocytic leukemia. In addition, the pathogenesis, diagnosis, and treatment of chylothorax are reviewed. PMID- 15043341 TI - Isolated splenic metastasis from primary lung adenocarcinoma. AB - Metastatic disease involving the spleen is uncommon, and isolated metastasis to the spleen is extremely rare. Most patients with splenic metastases have widely disseminated metastatic disease. A current literature review shows the incidence of isolated splenic metastasis ranges from 0 to 26% of all patients with splenic metastases. The reported primary malignancies of patients with splenic metastases include lung, colorectal, endometrial, ovarian, thyroid, pancreatic, gastric cancers, and, most commonly, melanoma. Although most patients with splenic metastases are clinically asymptomatic for splenic lesions, there have been reports of painful splenomegaly, splenic vein thrombosis, and splenic rupture, making diagnosis and consideration of prompt therapeutic intervention important. The time from diagnosis of a primary lung tumor to the discovery of splenic metastases ranges from 0 to 8 years in the literature. We report a rare case of an asymptomatic, isolated splenic metastasis in a 72-year-old man diagnosed 25 months after resection of an adenocarcinoma of the lung. PMID- 15043340 TI - Pernicious anemia: presentations mimicking acute leukemia. AB - Vitamin B12 deficiency can cause profound alterations in the bone marrow. These alterations can mimic the more serious diagnosis of acute leukemia. The two patients described in this report were originally suspected of having acute leukemia or myelodysplasia on the basis of the bone marrow smear, and induction chemotherapy was considered. However, after further studies, they were both found to have vitamin B12 deficiency, and parenteral vitamin B12 administration resulted in normalization of the bone marrow. PMID- 15043342 TI - Solitary recurrent metastasis of squamous cell carcinoma of the uterine cervix in the spleen: case report. AB - In cervical squamous cell carcinoma, solitary metastasis to and recurrence in the parenchyma of the spleen are uncommon in the absence of apparent disease in other sites. A case of a 50-year-old patient with a Stage IIa carcinoma of the cervix treated with radical hysterectomy and pelvic lymphadenectomy followed by radiotherapy is reported. Serial squamous cell carcinoma (SCC) antigen measurements have been performed for monitoring the course of disease, response to treatment, and detection of tumor recurrence. Recurrent disease was initially suspected on the basis of an elevated SCC level, 7.11 microg/ml (normal, < 1.5 microg/ml), subsequently confirmed by computed tomographic scan, magnetic resonance imaging, and positron emission tomography of the abdomen and pelvis. Laparoscopic splenectomy followed by chemotherapy with cisplatin was performed. The SCC antigen level was within normal limits postoperatively. The patient was well and clinically free of disease for 5 months. PMID- 15043343 TI - Atypical presentation of metastatic basal cell carcinoma. AB - Basal cell carcinoma is an indolent, slow-growing tumor that rarely metastasizes. Approximately 70% of tumors occur in the head and neck regions. If a basal cell tumor metastasizes, it usually spreads to the regional lymph nodes first, followed by the lungs. We describe a patient with basal cell carcinoma of the right lower extremity with skin metastases. Skin biopsy of one tumor revealed fibroepithelioma of Pinkus, a rare variant of basal cell carcinoma. PMID- 15043344 TI - Bilateral synchronous carcinoma of the male breast in a patient receiving estrogen therapy for carcinoma of the prostate: cause or coincidence? AB - An unusual case of bilateral, synchronous breast cancer in a male patient who had a history of long-term estrogen therapy for prostate cancer is presented. The possible role of estrogen in the development of male breast cancer is revisited. PMID- 15043345 TI - Anorectal melanoma: report of three cases with extended follow-up. AB - Primary anorectal melanoma is rare. There is controversy regarding the best surgical treatment because of its poor prognosis. Three cases with extended follow-up are reported in this article. A 53-year-old woman with rectal bleeding was diagnosed with a melanoma of the rectum and underwent an abdominoperineal resection. The patient died with distant metastases 8 months later. An 80-year old woman with rectal bleeding was diagnosed with a melanoma of the rectum and underwent a transanal local excision. She remains alive 4 years later but with locally recurrent disease. A 78-year-old man with rectal bleeding was diagnosed with a melanoma of the rectum and underwent an abdominoperineal resection. He died with local and metastatic disease 25 months later. Recent trends favor local excision when technically feasible, although some patients may require an abdominoperineal resection of the rectum, especially for larger tumors. PMID- 15043346 TI - Endobronchial myofibroblastic sarcoma presenting with hemoptysis in a patient with a normal chest x-ray. AB - Hemoptysis is an important symptom in clinical practice. The diagnosis of the underlying cause is often difficult, particularly in patients presenting with a normal chest x-ray. We report a case of hemoptysis with a normal chest x-ray due to a rare endobronchial neoplasm: myofibroblastic sarcoma. PMID- 15043347 TI - Renal actinomycosis mimicking renal tumor: case report. AB - A 68-year-old man was admitted after fever and general fatigue with severe inflammatory signs and anemia. T1- and T2-weighted magnetic resonance imaging showed low- to isointensity and low-intensity tumor in the right kidney, respectively, suggesting renal actinomycosis. However, the right kidney was explored transabdominally because the possibility of renal malignant lymphoma could not be excluded. After nephrectomy, characteristic colonies of Actinomyces were seen microscopically, and the histologic diagnosis was renal actinomycosis. The patient was treated with antibiotics and made good progress after operation. This case highlights the importance of magnetic resonance imaging for the diagnosis of renal actinomycosis. PMID- 15043348 TI - Anticonvulsant hypersensitivity syndrome: treatment with corticosteroids and intravenous immunoglobulin. AB - We describe the case of a patient in whom anticonvulsant hypersensitivity syndrome developed during treatment with phenytoin and progressed when therapy was changed to phenobarbital. Although therapeutic options remain controversial, corticosteroids and IV immunoglobulin were used in our patient. The patient had a complete recovery, suggesting the potential benefit of corticosteroids and IV immunoglobulin for anticonvulsant hypersensitivity syndrome. PMID- 15043349 TI - Nicotine replacement therapy-associated syndrome of inappropriate antidiuretic hormone. AB - Nicotine has been documented to regulate the release of plasma arginine vasopressin (AVP). The literature is inconclusive about the effects of nicotine replacement therapy on AVP release, although cigarette smoking has been shown to increase the release of AVP. No clinical case reports have documented the possible association between nicotine replacement and hyponatremia through AVP release. We report a case of a 39-year-old man who experienced syndrome of inappropriate antidiuretic hormone while on nicotine patch therapy. We theorize that the constant serum concentration of nicotine levels provided through the patch may cause hyponatremia through the continuous stimulation of vasopressin. PMID- 15043350 TI - Health-related quality of life and disability in survivors of multiple trauma one year after intensive care unit discharge. AB - OBJECTIVE: To evaluate health-related quality of life and disability in multiple trauma patients requiring intensive care unit management. DESIGN: A total of 87 survivors of multiple trauma, with a median age of 31 yrs and a median Injury Severity Score of 22, were enrolled in the present study. The Nottingham Health Profile, Glasgow Outcome Scale, and Rosser Disability Scale were used to assess the functional consequences of trauma 1 yr after intensive care unit discharge. RESULTS: A total of 64 of 87 patients had a problem in at least one of the six domains related to subjective health status. The most prevalent complaint was related to somatic subdimensions, but emotional functioning was also affected. Nottingham Health Profile part 2 showed that 63 of the survivors experienced problems in at least one of the daily activities. Of particular importance, inability to work was reported by 47% of the patients. Fifty-nine percent experienced moderate-to-severe disability as evaluated by Glasgow Outcome Scale and Rosser Disability Scale. High aggregate injury severity score along with severe head trauma were independent predictors of poor health-related quality of life and disability. CONCLUSIONS: The majority of survivors of major trauma exhibit considerable levels of disability and impairment in health-related quality of life. Global injury severity score and degree of brain trauma determine functional limitations. This information may help in organizing long term rehabilitation of multiple-trauma patients. PMID- 15043351 TI - Effects of a new spinal orthosis on posture, trunk strength, and quality of life in women with postmenopausal osteoporosis: a randomized trial. AB - OBJECTIVE: One fourth of women > or =50 yrs of age in the general population have one or more vertebral fractures. The orthotic treatment modality in the management of vertebral fractures caused by osteoporosis remains subjective because no objective data from clinical trials are available. The objective of this research was to evaluate the efficacy of a newly developed spinal orthosis in patients with osteoporotic vertebral fractures. DESIGN: We conducted a study that measured trunk muscle strength, angle of kyphosis, body height, body sway, and variables of quality of life such as pain, well-being, and limitations of daily living. RESULTS: Wearing the orthosis for 6-mo period was associated with a 73% increase in back extensor strength, a 58% increase in abdominal flexor strength, an 11% decrease in angle of kyphosis, a 25% decrease in body sway, a 7% increase in vital capacity, a 38% decrease in average pain, a 15% increase in well-being, and a 27% decrease in limitations of daily living. The overall tolerability of the orthosis was good, no side-effects were reported, and the drop-out rate of 3% was rather low. CONCLUSIONS: The use of an orthosis increases trunk muscle strength and thus improves posture in patients with vertebral fractures caused by osteoporosis. In addition, a better quality of life is achieved by pain reduction, decreased limitations of daily living, and improved well-being. Therefore, the use of an orthosis may represent an efficacious nonpharmacologic treatment option for spinal osteoporosis. PMID- 15043352 TI - Concurrent (tandem) cervical and lumbar spinal stenosis: a 10-yr review of 54 hospitalized patients. AB - OBJECTIVE: To determine the frequency of concurrent cervical and lumbar spinal stenosis (i.e., "tandem spinal stenosis" [TSS]) in a 10-yr review of hospital admissions. DESIGN: In a retrospective study of 460,964 hospital admissions to our 1,000-bed academic community hospital, we identified a base population of 54 patients with TSS. These patients presented with multiple complaints, including neurogenic claudication, progressive gait disturbances, and neurologic signs of both upper and lower motor neuron dysfunction, among others. Since 1964 when TSS was first described, estimates of its frequency have varied widely, ranging from a low of 5% to a high of 28%. RESULTS: Of the 54 cases of TSS identified among the 460,964 patient admissions during a 10-yr period, 36 were men and 18 were women. A total of 51 were >51 yrs of age. For all ages, the frequency rate of TSS in this series was 12 per 100,000 admissions. CONCLUSION: With TSS, the symptoms of either the cervical or the lumbar type initially predominate. Often, only after the primary pathology is treated does the secondary problem become evident. Although TSS occurred relatively infrequently in this series, its potential presence should not be overlooked. PMID- 15043353 TI - Cost-effectiveness of duplex ultrasound surveillance in spinal cord injury. AB - OBJECTIVE: To determine cost-effectiveness of surveillance with duplex ultrasound for thromboembolic disease in individuals with acute traumatic spinal cord injury at admission to rehabilitation. DESIGN: In this retrospective sequential case series study, individuals with traumatic spinal cord injury admitted to our rehabilitation facility between July 1, 1988, and December 31, 1998, were identified. Cost at our institution for treatment of thromboembolic disease was tabulated in 2001-2002 dollar amounts. Using this cost information, in a statistical model founded on available medical evidence, analysis for cost effectiveness was then performed. RESULTS: A total of 369 subjects met inclusion criteria. Based on the statistical model, the cost and estimated mortality from thromboembolic disease per subject when performing admission duplex ultrasounds was $449.28 and 0.016%, respectively. The estimated total cost per subject when not performing admission duplex ultrasounds was $136.29, with predicted mortality of 0.524% due to thromboembolic disease. Therefore, cost of one life saved when performing admission duplex ultrasound surveillance is calculated to be $61,542, with cost per life year gained between $1193 and $9050, depending on age at time of injury and severity of injury. CONCLUSION: Duplex ultrasound is a cost effective tool for deep venous thrombosis surveillance in individuals with acute traumatic spinal cord injury admitted to rehabilitation programs. PMID- 15043354 TI - Botulinum toxin type B in piriformis syndrome. AB - OBJECTIVE: To evaluate the clinical safety and efficacy of botulinum toxin type B (MYOBLOC) in reducing myofascial pain associated with piriformis syndrome. DESIGN: This was a single-center, outpatient, open-label study of patients with piriformis syndrome. Subjects were treated unilaterally or bilaterally, depending on their symptoms. Evaluations and procedures were performed by a single examiner who was not blinded, and there were no control subjects in this case series. Each piriformis muscle was infiltrated from one injection site under electromyographic guidance with 5000 units of botulinum toxin type B. RESULTS: A total of 20 patients were enrolled in this study. Significant reductions in mean visual analog scale scores for buttock and hip pain were noted at weeks 4, 12, and 16 and for low back pain at weeks 2, 12, and 16. Visual analog scale scores for general and low back pain, pain radiating into lower limbs, and tingling were significantly lower at week 2 after injection, suggesting early onset. A total of 95% of patients reported fair to excellent improvement in pain. Botulinum toxin type B was considered to have fair to excellent efficacy in 90% of patients, as evaluated by the investigator rating of overall efficacy. Botulinum toxin treatment was well tolerated. Dry mouth was the most common treatment emergent adverse event, reported in 6 of 20 patients. CONCLUSIONS: Findings suggest the possibility that botulinum toxin type B may be of potential benefit in the treatment of pain attributed to piriformis syndrome. PMID- 15043355 TI - PoWER Program: People with Disabilities Educating Residents. AB - An educational program was developed incorporating people with disabilities (PWDs) as "consumer-trainers" who educate physical medicine and rehabilitation residents about challenges in everyday living and their solutions to these. During the residency postgraduate years 2 and 4, pairs of residents visited trainers in their homes and at their jobs to learn about their lifestyles and adaptations. Residents' described the experience as "excellent, outstanding, invaluable" in more than half of the evaluations. PWDs can educate physical medicine and rehabilitation residents about living with disabilities. This new residency educational strategy encourages residents to value the perspective of PWDs. PMID- 15043356 TI - Brainstem auditory-evoked potentials as an objective tool for evaluating hearing dysfunction in traumatic brain injury. AB - Because of the violent nature of traumatic brain injury, traumatic brain injury patients are susceptible to various types of trauma involving the auditory system. We report a case of a 55-yr-old man who presented with communication problems after traumatic brain injury. Initial results from behavioral audiometry and Weber/Rinne tests were not reliable because of poor cooperation. He was transferred to our service for inpatient rehabilitation, where review of the initial head computed tomographic scan showed only left temporal bone fracture. Brainstem auditory-evoked potential was then performed to evaluate his hearing function. The results showed bilateral absence of auditory-evoked responses, which strongly suggested bilateral deafness. This finding led to a follow-up computed tomographic scan, with focus on bilateral temporal bones. A subtle transverse fracture of the right temporal bone was then detected, in addition to the left temporal bone fracture previously identified. Like children with hearing impairment, traumatic brain injury patients may not be able to verbalize their auditory deficits in a timely manner. If hearing loss is suspected in a patient who is unable to participate in traditional behavioral audiometric testing, brainstem auditory-evoked potential may be an option for evaluating hearing dysfunction. PMID- 15043357 TI - Efficacy of glossopharyngeal breathing for a ventilator-dependent, high-level tetraplegic patient after cervical cord tumor resection and tracheotomy. AB - This case study was undertaken to describe the use and limitations of glossopharyngeal breathing (GPB) by a ventilator-dependent, tracheotomized patient after cervical tumor resection. A 6-yr, 8-mo-old, tracheotomized, ventilator-dependent boy, after cervical tumor resection, learned GPB on his own and used it for ventilator-free breathing. Over the next 16 yrs, his GPB efficacy improved to the point that, with a vital capacity of 670 ml, his GPB maximum single-breath capacity increased to 3300 ml. This was limited by the fact that at 2.9 l of lung volume, air began to leak around the tracheostomy tube walls and out of the stoma. Still, GPB permitted up to 12 hrs/day of ventilator-free breathing. Measurements of assisted peak cough flow and GPB lung insufflations exceeding vital capacity are the main measures that demonstrate adequate tube fit to permit effective GPB in the presence of an indwelling tracheostomy tube. PMID- 15043359 TI - Iliopsoas weakness: a clinical sign of lumbar spinal stenosis. PMID- 15043358 TI - Spinal epidural hematoma associated with oral anticoagulation therapy. AB - Spontaneous spinal epidural hematoma is an uncommon cause of spinal cord compression. It may be associated with various causative factors, but in many patients, anticoagulation can be implicated. It is noteworthy that many of the reported cases were anticoagulated in the therapeutic range. Spontaneous spinal epidural hematoma should be suspected in any patient receiving anticoagulant agents who complains of local or referred spinal pain associated with limb weakness, sensory deficits, or urinary retention. Early diagnosis and treatment are very important for the functional recovery of the patient. Spinal magnetic resonance imaging is the most suitable neuroradiological method for early diagnosis. Although primary management is the surgical evacuation of the spinal epidural hematoma via laminectomy, rare cases in which the patient is improving rapidly and progressively could be treated conservatively. A 22-yr-old man with a spontaneous spinal epidural hematoma who was receiving warfarin treatment for a mechanical aortic valve is presented in this article. PMID- 15043360 TI - Beyond the fence line: corporate social responsibility. AB - The ability to demonstrate acceptable performance against the expectations of a CSR movement is part of the corporate license to operate in many locations. Because health and medical programs are core elements of these activities, numerous opportunities exist for physicians to contribute to health and prosperity in underdeveloped locations. Individuals who are involved in designing and administrating these activities need to maintain critical objectivity about the actual consequences and maintain a close dialog with the intended beneficiaries. The value of the CSR agenda is beginning a long empirical trial. PMID- 15043361 TI - Community health outreach program of the Chad-Cameroon petroleum development and pipeline project. AB - A critical appraisal has been presented of the CHOP for a large-scale energy infrastructure development project that was implemented in two of the world's poorest countries. The project is under close scrutiny from various independent monitoring groups, civil society organizations, and human rights groups. Reviewing the achievements and shortcomings permits the extraction of important lessons that will be critical for the future adoption of the CHOP in the current setting and for the implementation of additional CHOPs elsewhere in the developing world. The authors believe that the design must be flexible, efficient, and innovative so that a CHOP promptly can address pressing public health issues as they arise (eg, epidemic outbreak) and include the needs and demands of the concerned communities. An innovative feature of the current project is the high degree and mix of public-private partnerships. The project's CHOP also relies on partnerships. As elaborated elsewhere, public-private partnerships should be seen as a social experiment--they reveal promise but are not the solution for every problem. For this CHOP, the focus is on partnerships between a multinational consortium, government agencies, and international organizations. The partnerships also include civil society organizations for monitoring and evaluation and local NGOs designated for the implementation of the selected public health interventions within the CHOP. The governments and their respective health policies often form the umbrella under which the partnerships operate. With the increase in globalization, however, the importance and capacities of governments have diminished, and there is growing private-sector involvement. Private enterprise is seen as an efficient, innovative, pragmatic, and powerful means to achieve environmental and social sustainability. Experiences with the partnership configurations in the current CHOP are of importance for tackling grand challenges in global health by applying a systemic approach. Other innovations of the project in general, and the CHOP in particular, are the strong emphases on institutional-capacity building, integration, and sustainability. In countries like Chad and Cameroon, there are serious shortages of well-qualified health personnel. The CHOP described in this article provides leverage for initiating better healthcare that will reduce the high burden of disease in the developing world. Reducing mortality rates for infants and children younger than 5 years in sub-Saharan Africa requires massive scaling-up of malaria-control interventions (eg, large-scale distribution of ITNs to protect millions of African children), thereby approaching the Abuja targets (see Armstrong Schellenberg et al). The local NGOs that took a lead within the framework of the CHOP in the distribution of ITNs and accompanying health education messages can extend these activities to communities living outside the vicinity of the project area. Serious shortcomings of the current CHOP, consistently identified by the external monitoring groups, include the lack of a regional health plan, cumulative impact assessment, and provision of clean water and sanitation outside the narrowly defined project area. This point is of central importance, particularly for Chad, where access to clean water and improved sanitation facilities is low. Another limitation of the current CHOP is the insufficient amount of significance addressed to tuberculosis and the apparent lack of concerted control efforts against HIV infection, AIDS, and tuberculosis. These criticisms, however, must be balanced against the lack of clarity in international discourse about the proper extent of responsibility of the corporate sector for dealing with the health problems of countries in which they do business. In an elegant analysis, the environmental risk factor "unsafe water, sanitation and hygiene" was shown to be one of the major contributors to loss of healthy life, particularly in the developing world. Provision of clean water and sanitation is a key factor for sustainable control of schistosomiasis and soil-transmitted helminths. Reduction of helminth infections might have a beneficial effect on the HIV and AIDS pandemic. The question still remains: What is, or should be, the scope and limits of responsibility of the corporate sector in solving these problems? There is a critical need for the monitoring and evaluation of the long-term impact of a CHOP that develops in parallel with a large development project, emphasizing the broadest possible determinants of health and well-being. To become operational, it requires the establishment and running of a longitudinal demographic surveillance system in the area and in adjacent areas that are unlikely to be affected by the project. This approach, coupled with regular household surveys for in-depth appraisal of health-seeking and asset indices, is the most promising source of data for impact measurement of health, poverty, and equity-related issues. It will facilitate subtle monitoring and surveillance activities, fostering a truly systemic approach by inclusion of all stake holders on the basis of the existing but constantly evolving system. PMID- 15043362 TI - Vaccination strategies. AB - By nature of their long-term deployment in challenging areas of the world, expatriate employees and their dependents are at increased risk for a number of infectious diseases. Targeted immunization strategies may decrease this risk. Expatriate employees and their dependents should be immunized before deployment, and programs should be established to provide booster immunizations if the risk is ongoing. When considering work-force productivity, work-force infectivity, and the common good, careful consideration should be given to establishing at least a basic immunization program for in-country nationals and their dependents. PMID- 15043363 TI - HIV/AIDS prevention programs: methodologies and insights from the dynamic modeling literature. AB - Given the quickly changing landscapes of both global HIV infection and the HIV prevention literature, it has become increasingly difficult to identify or design an effective HIV prevention program. The authors discuss methods of selecting an appropriate program so that readers can critically evaluate the literature and create a suitable program for their unique setting. The intent of this article is to provide the reader with analytic modeling tools and methodology to search effectively the literature so that an appropriate and successful HIV prevention program can by selected by an organization or project team. PMID- 15043364 TI - Public health planning tools and compliance: food and water. AB - Overall food and water safety can be managed effectively through careful analysis of risks; implementation of site-specific food and water safety programs, including HACCP systems; careful assignment of responsibilities for implementation; and follow-up evaluation of effectiveness. PMID- 15043365 TI - Medical and trauma evacuations. AB - Evacuating ill or injured expatriates on international assignments from globally challenging locations is a potent source of concern for individuals who are involved before, during, and after overseas assignments. It is essential to minimize the risk for such evacuations and to make the evacuations proceed as smoothly and safely as possible. The first half of this article addresses the planning and processes that are required to make inevitable medical evacuations as safe and smooth as possible. The second half reviews the non-medical consequences that medical and security evacuations can have for employees, dependents, and employers. PMID- 15043366 TI - Distant diagnosis. AB - Effective communications are essential for any operation and for management of health-related problems. Telemedicine has altered profoundly the manner in which primary care physicians can interact with hospitals and specialists. As dedicated medical ICT systems evolve, medical care will be delivered to patients in remote locations in a more efficient and cost-effective way. Companies invariably have a vision that healthcare decisions benefiting patient care should be made anytime and anywhere in a model that facilitates care at the point of need. The development of telemedicine programs is inevitable as business globalization expands to more challenging cultural and geographic areas. PMID- 15043367 TI - Chronic diseases in globally challenging locations: assessment, impacts, and control. AB - A decision regarding work assignment in challenging environments often is made on an individual basis. The physician must consider (1) objective medical information (eg, CVD risk profile, results of exercise stress testing; see Fig. 6); (2) psychologic assessment, including potential career impacts of the expatriate assignment; and (3) availability of specialty medical facilities and expertise in the host country. Although these individual decisions are important, health professionals should not lose sight of the opportunity to approach chronic diseases on a broader basis by adopting a public health approach for the work site and the surrounding community. PMID- 15043368 TI - Malaria: control strategies, chemoprophylaxis, diagnosis, and treatment. AB - Malaria caused by Plasmodium falciparum continues to be the principal medical threat to travelers in tropical zones. Eighty percent of malaria cases occur in sub-Saharan Africa. Prevention capacity nears 100%. In the event of infection, early diagnosis and bitherapy (treatment that includes two different active molecules) provide a nonfatal outcome. The greatest risk occurs in the weeks after returning home. The history of malaria has shown that it is important to anticipate the evolving nature of the disease and to be able to respond to it in a timely fashion. The efficiency of preventive treatments should lead toward the goal of zero malaria cases when dealing with travelers; however, virtually all empiric experience indicates that this goal rarely is reached. PMID- 15043369 TI - Sexually transmitted infections: prevention and management. AB - In the early 21st century, STI and HIV have been linked inextricably. Although the focus of this article is STI, some discussion on the diagnosis and management of individuals with HIV infection is necessary. The history of HIV diagnosis in the workplace is checkered. The authors have seen cases of prospective workers being subjected to HIV testing without their knowledge as part of a pre employment medical examination. If the test came back positive, the men were told that they would not be employed without explanation. This approach is a breech of the human rights of the individual being tested and cannot be condoned. Any HIV testing must be done with the full and informed consent of the individual, with counseling given before and after testing to enable individuals with HIV infection to seek care and protect their families and to give individuals without HIV infection counseling on risk reduction. Men and women who present with an STI are at risk for HIV infection. With increasing options for management and secondary prevention, it is important to recognize people who are at risk. This identification should be done through HIV VCT. The location, funding, and supervision of VCT sites related to workplace populations should be a subject for serious debate. Although fears of mass layoffs after HIV testing largely have been unfounded, it is natural for workers to be fearful, unless there is a clearly articulated policy stating that the company observes and enforces nondiscriminatory practices. The workplace examples show that syndromic STI management, allied to comprehensive prevention programs, can have a genuine and measurable impact on STI prevalence. The potential interventions and partners are listed in Table 2. A community-based, randomized study in Tanzania showed that the institution of a well-managed STI syndromic management program can reduce HIV incidence by up to 40%, in the context of a rising HIV epidemic. Presumptive STI treatment for female sex workers (see Box 1) may prove useful as a short-term measure to reduce high STI prevalence rates while more sustainable preventive and curative services are established. The laboratory diagnosis of STIs remains problematic in the face of commonly available technologies of limited sensitivity and specificity and often substandard quality-assurance practices. For these and other reasons, syndromic management became the recommended strategy for treating STIs. The availability of rapid, accurate, and inexpensive diagnostics, especially for cervical infections for women, would alter management recommendations. Work is being done by the WHO and others to develop and assess low-cost diagnostics. Managing STIs and altering the behavior that leads to STIs are essential elements of any HIV prevention and management program. The issues surrounding a practical, compassionate, and comprehensive HIV program can be difficult. Numerous publications exist to help program managers navigate these issues and appropriately tailor a program to the needs of individual organizations. Some of these publications are listed in the next section. PMID- 15043370 TI - Respiratory infections: SARS and tuberculosis. AB - Positive-microscopy TB is a real issue for the respiratory diseases department of N'Djamena General Hospital, with a prevalence rate of 37 cases in 100 patients. The delay of diagnosis for positive-microscopy TB is excessive. Patients seem to be more responsible for this delay than the healthcare facilities. The factors that delay treatment of TB in N'Djamena are that (1) TB diagnosis in peripheral hospitals lengthens physician delay, and (2) seeking initial treatment outside conventional medicine lengthens patient delay. The following recommendations arise from this study: For the public health ministry: Reboost the national program against TB in Chad. Increase personnel in the respiratory diseases department at the N'Djamena General Hospital. Provide refresher TB education to care providers, stressing the importance of communication with the patient. For the national program against TB in Chad: Initiate a public information, education, and communication campaign about TB. Implement use of DOTS nationwide. For physicians increase communication with patients. PMID- 15043371 TI - Medical problems in the returning expatriate. AB - Expatriates are at risk for a number of infectious diseases for which short-term travelers generally are not at risk. Returning expatriates should undergo a detailed physical examination and a basic set of laboratory tests; these tests should be tailored to their specific history and exposures. Febrile patients with an appropriate exposure history must be evaluated for malaria; other potential diagnoses may be determined by incubation period, geographic exposure, and associated symptoms. When evaluating an ill returned expatriate with fever, it is important to exclude malaria, typhoid, leishmaniasis, brucellosis, tuberculosis, HIV infection, and syphilis. Gastrointestinal irregularities in expatriates may be caused by a number of infectious and noninfectious causes, including intestinal helminthiasis, strongyloidiasis, schistosomiasis, liver flukes, and amebiasis. Eosinophilia in returned expatriates often is associated with an infectious process and should be evaluated. Many infections associated with long term overseas deployment may include dermatologic manifestations, including filariasis and leishmaniasis. PMID- 15043372 TI - Stress in expatriates. AB - With economic globalization, the rate and effects of culture shock stress in expatriates and their families are expected to increase. The stressor may occur from the micro-environment, the macro-environment, or the mega-environment. The effects of stress are frequently due to predictable reactions such as insecurity, distress, and homesickness. These symptoms can progress to more severe psychosomatic symptoms, including depressive reactions and overt psychopathic behavior. Cross-cultural training programs are currently the best preventive strategy for culture shock and subsequent stress; however, the true efficacy of these programs is not well characterized. Substance abuse is probably a significant underlying problem for international business travelers; however, the true magnitude and effects of this issue are not well characterized in the expatriate community. There are substantial opportunities for additional medical research in these areas. PMID- 15043373 TI - Continuity of care: process or outcome? PMID- 15043374 TI - Defining and measuring interpersonal continuity of care. AB - BACKGROUND: In an effort to learn more about the importance of continuity of care to physicians and patients, I reviewed the medical literature on continuity of care to define interpersonal continuity and describe how it has been measured and studied. METHODS: A search of the MEDLINE database from 1966 through April 2002 was conducted to find articles focusing on the keyword "continuity of patient care," including all subheadings. Titles and abstracts of the resulting articles were screened to select articles focusing on interpersonal continuity in the physician-patient relationship or on the definition of continuity of care. These articles were systematically reviewed and analyzed for study method, measurement technique, and research theme. RESULTS: A total of 379 original articles were found that addressed any aspect of continuity as an attribute of general medical care. One hundred forty-two articles directly related to the definition of continuity or to the concept of interpersonal continuity in the physician-patient relationship. Although the available literature reflects little agreement on how to define continuity of care, it is best defined as a hierarchy of 3 dimensions; informational, longitudinal, and interpersonal continuity. Interpersonal continuity is of particular interest for primary care. Twenty-one measurement techniques have been defined to study continuity, many of which relate to visit patterns and concentration rather than the interpersonal nature of the continuity relationship. CONCLUSIONS: Future inquiry in family medicine should focus on better understanding the interpersonal dimension of continuity of care. PMID- 15043375 TI - Provider continuity in family medicine: does it make a difference for total health care costs? AB - BACKGROUND: International comparisons of health care systems have shown a relationship at the macro level between a well-structured primary health care plan and lower total health care costs. The objective of this study was to assess whether provider continuity with a family physician is related to lower health care costs using the individual patient as the unit of analysis. METHODS: We undertook a study of a stratified sample of patients (age, sex, region, insurance company) for which 2 cohorts were constructed based on the patients' utilization pattern of family medicine (provider continuity or not). Patient utilization patterns were observed for 2 years. The setting was the Belgian health care system. The participants were 4,134 members of the 2 largest health insurance companies in 2 regions (Aalst and Liege). The main outcome measures were the total health care costs of patients with and without provider continuity with a family physician, controlling for variables known to influence health care utilization (need factors, predisposing factors, enabling factors). RESULTS: Bivariate analyses showed that patients who were visiting the same family physician had a lower total cost for medical care. A multivariate linear regression showed that provider continuity with a family physician was one of the most important explanatory variables related to the total health care cost. CONCLUSIONS: Provider continuity with a family physician is related to lower total health care costs. This finding brings evidence to the debate on the importance of structured primary health care (with high continuity for family practice) for a cost-effective health policy. PMID- 15043376 TI - Continuity of primary care: to whom does it matter and when? AB - BACKGROUND: Inconsistent findings on the value of continuity of care can stem from variability in its importance to different subsets of patients. We therefore examined the association among patient and visit characteristics and extent to which the patient valued continuity of care (PVC). We hypothesized that continuity would be more important to patients who are older, sicker, and female, who have established a relationship with their physician, and whose visit addresses more complex problems. METHODS: A study of 4,454 consecutive outpatient visits to 138 community-based family physicians used a 3-item measure (alpha = 0.67) of PVC. The patient's report of (1) the adequacy of primary care for the visit and (2) satisfaction with the physician on that visit was assessed with multiple measures. Analyses examined the associations among PVC and patient reported satisfaction with the physician and adequacy of the visit. RESULTS: Extremes of age, female sex, less education, Medicare and Medicaid insurance, number of chronic conditions and medications, number of visits to the practice, and worse self-reported health status were associated with higher value placed on continuity (P < .001 for all except sex, where P = .015). Patients who value continuity and did not see a regular physician rated adequacy of the visit lower (for 7 attributes of the visit) than those seeing their own physician. Satisfaction with the physician for the visit was greatest among patients who value continuity and saw their regular physician. CONCLUSIONS: Continuity of physician care is associated with more positive assessments of the visit and appears to be particularly important for more vulnerable patients. Health care systems and primary care practices should devote additional effort to maintaining a continuity relationship with these vulnerable patients. PMID- 15043377 TI - On being new to an insurance plan: health care use associated with the first years in a health insurance plan. AB - BACKGROUND: We wanted to compare health care utilization and costs in the first year of being in a health insurance plan with those of subsequent years. METHODS: We used claims data from an independent practitioner association (IPA)-style managed care organization in the Rochester, NY, metropolitan area from 1996 through 1999. Cross-sectional and panel analyses of up to 4 years of claims data were conducted, involving 335,547 adult patients assigned to the panels of 687 primary care physicians (internists and family physicians). Multivariate analyses, adjusting for age, sex, case mix, and socioeconomic status derived from ZIP codes, examined the relationship between the first year of health insurance and Papanicolaou tests, mammograms in women older than 40 years, physician use, avoidable hospitalization, and expenditures. RESULTS: After multivariate adjustment, the first year of insurance was associated with a higher risk of not getting a mammogram, a higher risk of avoidable hospitalization, greater likelihood of visiting a physician, and higher expenditures, especially for testing. There was no relationship, however, between Papanicolaou test compliance and year of enrollment. CONCLUSIONS: The findings suggest there might be adverse clinical and financial implications associated with changing insurance. PMID- 15043379 TI - Who first suggests the diagnosis of attention-deficit/hyperactivity disorder? AB - PURPOSE: We wanted to survey physicians in the Washington, DC, metropolitan area regarding the frequency with which physicians, teachers, parents, and others first suggest the diagnosis of attention-deficit/hyperactivity disorder (ADHD). METHODS: A questionnaire was mailed to all family physicians, primary care pediatricians, and child psychiatrists in greater Washington, DC. In answer to the question, "Who first suggests the diagnosis of ADHD?" respondents assigned percentages to primary care physicians, consultants, parents, teachers, etc. RESULTS: Teachers were most likely to be first to suggest the diagnosis of ADHD (46.4%; 95% CI, 44.1%-48.7%), followed by parents (30.2%; 95% CI, 28.3%-32.0%), primary care physicians (11.3%; 95% CI, 9.7%-12.8%), school personnel other than teachers (6.0%; 95% CI, 4.9%-7.2%), consultants such as child psychiatrists or psychologists (3.1%; 95% CI, 2.3%-3.9%) and other specified categories (3.0%; 95% CI, 2.4%-3.6%). CONCLUSIONS: Teachers and other school personnel are often the first to suggest the diagnosis of ADHD in children in the greater Washington, DC, area. Regional variations in the prescribing of medication for ADHD may be caused at least in part by variations in the likelihood of a teacher suggesting the diagnosis of ADHD. PMID- 15043380 TI - A journey to someplace better. PMID- 15043381 TI - Long-time researcher Paul Nutting wins 2003 Curtis G. Hames Research Award. PMID- 15043378 TI - Impact of provider continuity on quality of care for persons with diabetes mellitus. AB - BACKGROUND: Many patients with diabetes fail to receive recommended monitoring tests. One reason might be inadequate continuity of care. This study examined the association between provider continuity and completion of monitoring tests for patients with diabetes mellitus. METHODS: A cross-sectional analysis was conducted on claims data from a private national health plan for 1 year (January 1, 1999, through December 31, 1999). Participants had a diagnosis of diabetes mellitus and at least 2 outpatient visits during the study year (N = 1,795). The association was measured between continuity of care with an individual provider and completion of 3 diabetes monitoring tests: a glycosylated hemoglobin test, a lipid profile, and an eye examination. RESULTS: Eighty-one percent of patients had a glycosylated hemoglobin test, 66% had a lipid profile, and 28% had an eye examination during the study year. After controlling for demographics, number of diabetes visits, case mix, and diabetes complications, provider continuity was not significantly associated with the receipt of a glycosylated hemoglobin test (odds ratio [OR] = 0.61, 95% confidence interval [CI], 0.32-1.16), a lipid profile (OR = 0.97, 95% CI, 0.57-1.64) or an eye examination (OR = 0.60, 95% CI, 0.30-1.19). When continuity was measured only among primary care providers, there was no significant association for receipt of a glycosylated hemoglobin test (OR = 0.73, 95% CI, 0.41-1.33), a lipid profile (OR = 0.88, 95% CI, 0.53-1.47) or an eye examination (OR = 0.70, 95% CI, 0.35-1.36). CONCLUSIONS: This study found no association between provider continuity and completion of diabetes monitoring tests in a national privately insured population. Whereas continuity might benefit other aspects of health care, it does not appear to benefit improved monitoring for diabetes. PMID- 15043382 TI - Balanced scorecards: a new tool for family medicine. PMID- 15043383 TI - Community needs assessment and development using the participatory research model. PMID- 15043384 TI - Advancing health care coverage for all. PMID- 15043385 TI - The increasing problem of cephalosporin resistance mediated by extended-spectrum beta-lactamases. PMID- 15043386 TI - Current therapy and the development of therapeutic options for the treatment of diseases due to bacterial agents of potential biowarfare and bioterrorism. AB - An important part of biodefense is the optimization of current therapy and the development of new therapeutic options for the treatment of the diseases most likely encountered in the form of biological weapons. Guidelines for the prevention and treatment of anthrax, plague, tularemia and botulinum toxin intoxication are reviewed. The strategies in development for the prevention of anthrax focus primarily on active and passive immunization against protective antigen, because of its central role as a toxin delivery module. Novel vaccine strategies for plague, tularemia and botulism are also reviewed. PMID- 15043387 TI - CpG DNA as a potent inducer of mucosal immunity: implications for immunoprophylaxis and immunotherapy of mucosal infections. AB - Recent advances in immunology reveal that vertebrate innate immune systems use pattern-recognition receptors, of which the best characterized is the toll-like receptor (TLR) family, to specifically detect pathogen-associated molecular patterns (PAMPs) present in infectious agents. Based on this, use of synthetic PAMPs has attracted much interest for specific immunoprophylaxis and immunotherapy purposes. Among the PAMPs, immunostimulatory CpG DNA, a TLR9 ligand, displays promising potent immunostimulatory and adjuvanticity in different settings. In this review, recent developments in the use of CpG DNA for inducing immunity in the mucosal tissues will be highlighted. PMID- 15043389 TI - Isoprenoid biosynthesis as a novel target for antibacterial and antiparasitic drugs. AB - The mevalonate-independent methylerythritol phosphate pathway is a long overlooked metabolic pathway for isoprenoid biosynthesis. It is present in most bacteria, including pathogens and opportunistic pathogens, in some unicellular eukaryotes, including the parasite responsible for malaria, and in the chloroplasts of all phototrophic organisms. It represents an alternative to the mevalonate pathway, which is only present in animals, fungi, the plant cytoplasm, archaebacteria and some eubacteria. This biosynthetic pathway is thus a potential target for antibacterial and antiparasitic drugs. An isopentenyl diphosphate isomerase that differs from the previously known isopentenyl diphosphate isomerase found in all other organisms, including animals, was discovered in several Gram-positive bacteria possessing the mevalonate pathway, adding another target related to isoprenoid biosynthesis. PMID- 15043390 TI - Advances in respiratory syncytial virus vaccine development. AB - Respiratory syncytial virus (RSV) is the most important causative agent of respiratory tract infections. Young children with chronic lung or congenital heart disease are at increased risk for severe disease. Intensive research into a candidate vaccine has yielded live attenuated vaccines and subunit vaccines, which have been studied in humans. Although immunogenic, occurrence of upper respiratory tract infection symptoms with live attenuated vaccine prohibits evaluation in young infants. Subunit vaccines include purified F protein (PFP-1 and -2) and BBG2Na. PFP vaccines are effective in seropositive children, but also induce upper respiratory symptoms. BBG2Na was being investigated in phase III clinical trials, however, further development has now been discontinued. This review discusses recent advances in RSV vaccine development. PMID- 15043388 TI - Fatty acid biosynthesis as a target for novel antibacterials. AB - The bacterial fatty acid synthesis pathway has significant potential as a target for the development of novel antibacterials. The pathway has been extensively studied in Escherichia coli, the crystal structures of the compounds involved are known and homologous genes are readily identified in the genomes of important pathogens. The currently used drugs triclosan and isoniazid are known to target one step in the pathway. Other experimental compounds such as thiolactomycin and cerulenin effectively inhibit other steps. These known pathway inhibitors are reviewed and the areas for potential future developments are explored. PMID- 15043391 TI - Novel treatment options for hepatitis B virus infection. AB - Chronic hepatitis B virus (HBV) infection affects between 350 and 400 million people globally. Interferon-alpha, lamivudine and adefovir (Hepsera) are approved for the treatment of chronic hepatitis B; however, the use of interferon-alpha is limited. Lamivudine and adefovir have excellent antiviral activity and oral bioavailability, although viral rebound after cessation of therapy and development of resistance after long-term therapy with lamivudine are major clinical limitations. Adefovir has proven to be effective against lamivudine resistant strains in vitro and in vivo. The various steps of HBV replication provide opportunities for new antiviral drugs to interact with the virus. Recent developments, including new antivirals that interfere with viral DNA replication, hold promise for the future. Sustained reduction in viral load and improved treatment of chronic HBV infection could, in future, be achieved by the development of new drugs with different mechanisms of action and resistance profiles, and with combination therapy involving two or more nucleosides with or without immunomodulators. PMID- 15043392 TI - Severe acute respiratory syndrome: scientific and anecdotal evidence for drug treatment. AB - Severe acute respiratory syndrome (SARS), caused by a highly infectious novel coronavirus (CoV), predominantly presents with severe pneumonitis leading to respiratory failure and death in approximately 10% of victims. Most cases present, after an incubation of 2 to 11 days, with fever and chills, which are followed by dry cough and dyspnea before the onset of respiratory failure. The management of SARS is controversial, largely due to the lack of data from controlled trials, which were logistically impossible to design or execute at the time of the overwhelming outbreak between March and June 2003. The use of an antiviral is logical although there is no effective agent against SARS-CoV, with the widespread use of ribavirin in 2003 attracting considerable scepticism. The use of ribavirin as a monotherapy in SARS is not recommended. Retrospective data suggest that administration of the anti-HIV drug Kaletra in combination with ribavirin could reduce mortality and incidence of respiratory failure. The use of corticosteroid was based on the similarity between SARS and bronchiolitis obliterans organizing pneumonia. Corticosteroid use is considered important in subsets of patients with SARS. As SARS may still resurge, it is imperative that past experience is analyzed. This review will attempt to address the rationale for pharmacotherapy in SARS using anecdotal and the limited published data. PMID- 15043393 TI - Candida biofilms: antifungal resistance and emerging therapeutic options. AB - Intravascular catheter infections are a major cause of morbidity and mortality in hospitalized patients, accounting for the majority of the 200,000 nosocomial bloodstream infections occurring in the US annually. Of the intravenous lines that are culture-positive for Candida, 40% actually represent fungemia, which generally necessitates systemic treatment and line removal to affect cure. Until recently, the reason for the need for device removal was unclear. However, our research group and others have demonstrated a near-total resistance to antifungals by biofilm-associated Candida. Similar to bacterial species, Candida biofilm formation proceeds through early, intermediate and maturation phases. This process is associated with the generation of a polysaccharide extracellular matrix (ECM). Mature C. albicans biofilms have a heterogeneous architecture, in terms of distribution of fungal cells and ECM, and exhibit broad antimicrobial resistance. The mechanisms causing such profound antifungal resistance are beginning to be understood. Recent data indicate that resistance is phase specific and multifactorial, involving efflux pumps and sterol synthesis (at early and mature biofilm phases, respectively). Neither metabolic quiescence nor the ECM appear to contribute substantially. Susceptibility testing and confocal scanning laser microscopy demonstrated that azoles failed to exert activity against mature Candida biofilms. However, sub-inhibitory concentrations of voriconazole impaired biofilm formation and caused cell morphological aberrations. In contrast, lipid-formulation amphotericins and the echinocandins uniquely exhibited activity against mature biofilms. The mechanisms underlying this ability are unknown. The role of other pharmacological (eg, catheter coatings, antimicrobial peptides and antibiotic locks) and non-pharmacological methods in the prevention and treatment of device-related biofilms is discussed in this review. PMID- 15043394 TI - Amphotericin B cochleates: a vehicle for oral delivery. AB - Cochleates are a novel lipid-based delivery vehicle consisting of crystalline phospholipid-cation structures that form spiral lipid sheets. They represent a new technology platform for oral delivery of clinically important drugs that possess poor oral bioavailability. Orally administered cochleates containing amphotericin B (CAMB) showed broad-spectrum activity in murine infection models of candidiasis, aspergillosis and cryptococcosis. Initial biodistribution studies of CAMB administered orally in mice demonstrated that cochleates delivered significant levels of AMB to target organs. The lipid particulate nature of cochleates also imparted reduced toxicity that mimics other lipid-amphotericin B complexes. Cochleates are a promising new vehicle for oral delivery of amphotericin B at therapeutic levels. PMID- 15043395 TI - Antifungal therapy for febrile neutropenia: issues in clinical trial design. AB - Amphotericin B therapy for fever during neutropenia became a standard practice approximately 20 years ago. Treatment of febrile neutropenia has since become a common indication sought in the development of new antifungals, with several alternatives approved for 'empirical therapy' based on the results of randomized clinical trials. Recent studies have highlighted problematic issues in trial design and interpretation; inclusion criteria and the definition of success are two examples of controversial issues. Here, issues in trial design will be reviewed to address the question: Can (and should) new antifungals be evaluated for treatment of fever during neutropenia? PMID- 15043396 TI - TMC-125 Tibotec. AB - TMC-125, a non-nucleoside reverse transcriptase inhibitor, is being developed by Tibotec for the potential treatment of HIV infection. Phase IIa trials in treatment-naive and treatment-experienced HIV-1-infected individuals had been completed by February 2002. A long-term, phase IIb, dose-finding study in treatment-experienced patients started enroling patients in a number of European countries during 2002, and Canada during 2003. PMID- 15043397 TI - AIDSVAX VaxGen. AB - AIDSVAX, a bivalent vaccine consisting of a preparation of recombinant gp120 from two types of HIV, is being developed by VaxGen for the potential prevention of HIV infection. Two versions of the vaccine are in phase III trials: AIDSVAX B/E, in trials in Thailand, stimulates production of antibody to HIV subtypes common to South Asia, South East Asia and countries of the Pacific Rim; and AIDSVAX B/B, in trials in the US and Europe, stimulates the production of antibody to HIV subtypes that are found in the Americas, Europe, the Caribbean and Australia. PMID- 15043398 TI - Glyminox Biosyn. AB - Glyminox (Savvy) is a vaginal gel formulation of C31G being developed by Biosyn as a potential contraceptive and for the potential prevention of transmission of sexually transmitted diseases, including HIV infection. In March 2002, glyminox was in phase II/III trials for Chlamydia treatment and as a contraceptive, and in phase II trials for herpes simplex virus shedding. PMID- 15043399 TI - Telbivudine/Torcitabine Idenix/Novartis. AB - Idenix (formerly Novirio) and Novartis are developing two beta-L deoxynucleosides, telbivudine and torcitabine, for the potential treatment of hepatitis B virus infection. Phase III trials of telbivudine were underway by August 2002 and phase I/II trials of the torcitabine prodrug, valtorcitabine, were ongoing in November 2003. PMID- 15043400 TI - Manganese toxicokinetics at the blood-brain barrier. AB - Increased manganese (Mn) use in manufacturing and in gasoline has raised concern about Mn-induced parkinsonism. Previous research indicated carrier-mediated brain entry but did not assess brain efflux. Using in situ rat brain perfusion, we studied influx across the blood-brain barrier (BBB*) of three predominant plasma Mn species available to enter the brain: Mn2+, Mn citrate, and Mn transferrin. Our results suggested transporter-mediated uptake of these species. The uptake rate was greatest for Mn citrate. Our results using the brain efflux index method suggested that diffusion mediates distribution from rat brain to blood. To characterize the carriers mediating brain Mn uptake, we used rat erythrocytes, an immortalized murine BBB cell line (b.End5), primary bovine brain endothelial cells (bBMECs), and Sprague Dawley and Belgrade rats. Studies with bBMECs and b.End5 cells suggested concentrative brain Mn2+ and Mn citrate uptake, respectively, consistent with carrier-mediated uptake. Mn2+ uptake positively correlated with pH, suggesting mediation by an electromotive force. Mn2+ uptake was not inhibited by iron or the absence of divalent metal transporter 1 (DMT-1) expression, suggesting an iron-transporter-independent mechanism. Mn2+ uptake inversely correlated with calcium and was affected by calcium channel modulators, suggesting a role for calcium channels. Rat erythrocyte results suggested monocarboxylate transporter 1 (MCT1) and anion exchange transporters do not mediate Mn citrate brain uptake. Considering carrier-mediated brain influx (but not efflux), repeated excessive Mn exposure should produce brain accumulation. Further work is necessary to identify the specific transporter or transporters mediating Mn distribution across the BBB. PMID- 15043401 TI - Petrosifungins A and B, novel cyclodepsipeptides from a sponge-derived strain of Penicillium brevicompactum. AB - A strain of Penicillium brevicompactum derived from a specimen of the Mediterranean sponge Petrosia ficiformis was investigated for its secondary metabolites. Using fast centrifugal partitioning chromatography (FCPC) two previously unknown cyclodepsipeptides, petrosifungins A (1) and B (2), were isolated, both containing two neighboring units of the nonproteinogenic amino acid l-pipecolinic acid. The absolute configurations of all amino acids were established to be l using GITC derivatization and Marfey's method. Furthermore, the known metabolite brevianamide A (3) was isolated. The likewise known compounds mycophenolic acid (4) and asperphenamate (5) were identified from their spectroscopic properties directly in the extract using HPLC-UV, -MS, and -NMR coupling. PMID- 15043402 TI - New cytotoxic clerodane diterpenoids from the leaves and twigs of Casearia membranacea. AB - Bioassay-guided fractionation of an EtOAc-soluble extract of Casearia membranacea has resulted in the isolation of six new clerodane diterpenes, caseamembrins A-F (1-6), and a known compound, rel-(2S,5R,6R,8S,9S,10R,18S,19R)-diacetoxy-18,19 epoxy-6-hydroxy-2-(2-methylbutanoyloxy)cleroda-3,13(16),14-triene (7). The structures of 1-6 were established on the basis of extensive 1D and 2D NMR spectroscopic analysis. In addition, the new derivatives, 8 and 9, were prepared by acylation of 7 and 3, respectively. The isolated diterpenoids and their derivatives were tested against human prostate (PC-3) and hepatoma (Hep3B) cancer cells. Compounds 1, 3-5, and 7 exhibited cytotoxicity against both tumor cells, with IC(50) values below 3 micromicro, while compounds 2, 6, 8, and 9 were less effective. PMID- 15043403 TI - Antineoplastic agents. 511. Direct phosphorylation of phenpanstatin and pancratistatin. AB - Selective phosphorylation of phenpanstatin (3a) with tetrabutylammonium dihydrogen phosphate and dicyclohexylcarbodiimide in pyridine followed by cation exchange chromatographic procedures was found to provide an efficient route to a new series (3b-3d) of promising 3,4-O-cyclic phosphate prodrugs designated phenpanstatin phosphates. Application of analogous reaction conditions to pancratistatin (1a) led to a mixture of monophosphate derivatives where sodium pancratistatin 4-O-phosphate (4a) was isolated and the structure confirmed by X ray crystallography. Modification of the reaction conditions allowed direct phosphorylation of pancratistatin followed by cation-exchange chromatography to afford sodium pancratistatin 3,4-O-cyclic phosphate (5a), which was selected for preclinical development. PMID- 15043404 TI - Aspochalasins I, J, and K: three new cytotoxic cytochalasans of Aspergillus flavipes from the rhizosphere of Ericameria laricifolia of the Sonoran Desert. AB - Bioassay-guided fractionation of a cytotoxic EtOAc extract of Aspergillus flavipes occurring in the rhizosphere of Ericameria laricifolia resulted in the isolation of three new cytochalasans, namely, aspochalasins I (1), J (2), and K (3), and four known cytochalasans, aspochalasins C (4), D (5), and E (6) and TMC 169 (7). The structures of compounds 1-3 were established on the basis of extensive 1D and 2D NMR spectroscopic analysis. All compounds exhibited weak to moderate cytotoxicity against NCI-H460, MCF-7, and SF-268 cancer cell lines, but none showed significant selectivity. PMID- 15043405 TI - New diterpenoids from Viburnum awabuki. AB - Eight new vibsane-type diterpenoids, vibsanins P-W (1-8), were isolated from the methanol extracts of the leaves and twigs of Viburnum awabuki. The structures were elucidated by 1D and 2D NMR spectral analysis, and their cytotoxicity against selected cancer cells was measured in vitro. PMID- 15043406 TI - Seco[D-Asp3]microcystin-RR and [D-Asp3,D-Glu(OMe)6]microcystin-RR, two new microcystins from a toxic water bloom of the cyanobacterium planktothrixrubescens. AB - Two novel microcystins, seco[d-Asp(3)]microcystin-RR (1) and [d-Asp(3),d Glu(OMe)(6)]-microcystin-RR (2), along with the known [d-Asp(3)]microcystin-RR (3), were isolated from a Planktothrix rubescens toxic bloom collected in Lake Bled, Slovenia. The structures were deduced using one- and two-dimensional NMR techniques, ESIMS/CID/MS analysis, and Marfey's method for determining the amino acids' absolute stereochemistry. Compounds 1 and 3 exhibit weak PP1 inhibitory activity. The NMR data of compound 3 are reported here for the first time. PMID- 15043407 TI - Cytotoxic constituents of the twigs and leaves of Aglaia rubiginosa. AB - Activity-guided fractionation of a CHCl(3)-soluble extract of the twigs of Aglaia rubiginosa, using human oral epidermoid carcinoma (KB) cells as a monitor, led to the isolation of a new naturally occurring cyclopenta[b]benzofuran, 1-O acetylrocaglaol (1), along with seven known compounds, methyl rocaglate (2), rocagloic acid (3), 1-O-acetylmethyl rocaglate (4), desyclamide, eryodictiol, 5 hydroxy-3,7,4'-trimethoxyflavone, and naringenin. A CHCl(3) extract of the leaves of A. rubiginosa yielded the new compound (3S,4R,22R)-cholest-7,24-diene-3,4,22 triol (5), as well as 11 known compounds, including 2 and 4 and cabraleone, dammarelonic acid, (20S,23E)-20,25-dihydroxy-3,4-secodammara-4(28),23-dienoic acid, (20S,23E)-20,25-dihydroxy-3,4-secodammara-4(28),23-dienoic acid methyl ester, (3beta,4beta,22R)-ergosta-5,24(24')-diene-3,4,22-triol, ocotillone, shoreic acid, beta-sitosterol, and beta-sitosterol glycoside. The structures of 1 and 5 were elucidated by spectral and chemical methods. Isolates were evaluated with a human cancer cell panel, and compounds 1-4 were found to exhibit potent cytotoxic activity. PMID- 15043408 TI - Effect of phenolic glycosides on Agrobacterium tumefaciens virH gene induction and plant transformation. AB - O-Aryl-d-glucoside (4-7) and d-xyloside (8-10) derivatives were synthesized and tested on Agrobacterium virH gene induction and plant transformation. alpha- or beta-Glycosides enhanced vir activity at concentrations above 250 micromicro. The highest vir activity was observed with beta-glucoside derivative 4 at 10 mM. A marked difference between phenol glucoside derivative 4 and the corresponding free phenol on the growth of transformants was observed. The regenerated transgenic tissues, after transformation on medium containing acetosyringyl beta glucoside 4, grew at twice the rate of those on medium containing only free acetosyringone (AS). Compound 4 was less toxic for tobacco explants compared to the corresponding free phenol. However, the xyloside derivatives tested (8-10) were less effective for gene induction compared with corresponding free phenols. PMID- 15043409 TI - Anthrone and oxanthrone C-glycosides from Picramnia latifolia collected in Peru. AB - Cytotoxicity-based, bioassay-guided fractionation of the chloroform-soluble extracts of both the roots and leaves of Picramnia latifolia led to the isolation of two new anthrone C-glycosides, picramniosides G (1) and H (2), two new oxanthrone C-glycosides, mayosides D (3) and E (4), and a new benzanthrone natural product, 6,8-dihydroxy-10-methyl-7H-benz[de]anthracen-7-one (5), together with 10 known compounds, 6,8-dihydroxy-4-methyl-7H-benz[de]anthracen-7-one (6), nataloe-emodin (7), chrysophanein, chrysophanol, 1,5-dihydroxy-7-methoxy-3 methylanthraquinone, pulmatin, 7-hydroxycoumarin, 7-hydroxy-6-methoxycoumarin, beta-sitosterol, and beta-sitosterol glucoside. The structures of 1-5 were established by spectroscopic methods, including 1D and 2D NMR, HRMS, and CD data interpretation. The cytotoxic activity of all isolates was evaluated in a small panel of human cancer cell lines. Compound 7 exhibited significant in vitro cytotoxic activity in the tested cell lines, but no significant activity was observed with an in vivo hollow fiber model at doses of 6.25, 12.5, 25, and 50 mg/kg/injection. PMID- 15043410 TI - Iridoids as allelochemicals and DNA polymerase inhibitors. AB - Growth inhibitory activities and nutritional indices of catalpol (1), 8-O acetylharpagide (2), and harpagide (3) were determinated in larvae and adults of Tribolium castaneum, respectively. Compound 1 produced a series of allelochemical effects probably related with the DNA synthesis. This iridoid possessed the highest inhibitory activity against DNA polymerase. Molecular orbital calculations suggest that a pi-pi charge transfer recognition model could explain the action of iridioids toward nucleic acid synthesis. PMID- 15043411 TI - Expanding the strategies in natural product studies of marine-derived fungi: a chemical investigation of penicillium obtained from deep water sediment. AB - Three previously unknown pentaketides, (+)-formylanserinone B (3), (-) epoxyserinone A (4), and (+)-epoxyserinone B (5), along with two known fungal pigments, anserinones A (1) and B (2), were isolated and identified from a deep water (-4380 ft), marine-derived saltwater fungal culture. Two other minor constituents, hydroxymethylanserinone B (6) and deoxyanserinone B (7), were also isolated, but not completely purified. The structures of 3-7, each expanding the dense functionalization of the anserinones, were determined by dereplication and spectroscopic analysis. Bioactivity was explored in two separate cell-based assays. Leukemia selectivity was greatest with 2 and 3, while 1-3 exhibited modest activity against the MDA-MB-435 cell line. PMID- 15043412 TI - Antifouling activity of brominated cyclopeptides from the marine sponge Geodia barretti. AB - In this work, we show the potent antifouling effects of two compounds, barettin (cyclo[(6-bromo-8-entryptophan)arginine]) (1), isolated as a Z/E mixture (87/13), and 8,9-dihydrobarettin (cyclo[(6-bromotryptophan)arginine]) (2), isolated from the marine sponge Geodia barretti. The compounds were isolated guided by their ability to inhibit the settlement of cyprid larvae of the barnacle Balanusimprovisus, and their structures were determined by means of mass spectrometry, NMR, and quantitative amino acid analysis. The activities of these brominated diketopiperazine-like cyclic dipeptides are in the range of antifouling agents in use today, as shown by their EC(50) values of 0.9 and 7.9 microM, respectively. However, contrary to today's antifouling agents, the effects of barettin and 8,9-dihydrobarettin are nontoxic and reversible. A small set of synthetic analogues, including l-arginine, l-tryptophan, 5-bromo-d,l tryptophan, 6-bromo-d,l-tryptophan, and 6-fluoro-d,l-tryptophan, were tested for possible structure-activity relationships. None of these compounds showed any effect at a concentration of 10 microM. We hypothesize that the isolated compounds are part of the sponge's chemical defense to deter fouling organisms. This theory is supported by the fact that barettin is found in water exposed to living specimens of G. barretti in concentrations that completely inhibit barnacles from settling. PMID- 15043413 TI - Excisanin H, a novel cytotoxic 14,20-epoxy-ent-kaurene diterpenoid, and three new ent-kaurene diterpenoids from Rabdosia excisa. AB - A novel 14,20-epoxy-ent-kaurene diterpenoid, excisanin H (1), and three new ent kaurene diterpenoids, 2-4, were isolated from aerial parts of Rabdosia excisa along with eight known ent-kaurene diterpenoids, 5-12. The structural elucidations were made using spectral (HREIMS, IR, (1)H, (13)C, and 2D NMR) methods. The absolute configuration of 1 was determined by demonstrating that oxidation of kamebakaurin (8) produced excisanin H (1). These ent-kaurene diterpenoids all showed significant cytotoxic activity against P388 murine leukemia cells. PMID- 15043414 TI - Oleanane saponins from Sanicula elata var. chinensis. AB - Eleven new oleanane-type triterpenoid saponins, saniculasaponins I-XI (1-11), and a known saponin, sandrosaponin IX (12), were isolated from the methanol extract of the whole plants of Sanicula elataHam. var. chinensis Makino. The structures of the new compounds were elucidated on the basis of chemical and spectroscopic evidence. PMID- 15043415 TI - Hydrocotylosides I-VII, new oleanane saponins from hydrocotylesibthorpioides. AB - Seven new oleanane-type triterpenoid saponins, hydrocotylosides I-VII (1-7), and one known saponin, udosaponin B (8), were isolated from the methanol extract of the whole plants of Hydrocotyle sibthorpioides.The structures of the new compounds were elucidated on the basis of chemical and spectroscopic evidence. PMID- 15043416 TI - New compounds from an extract of Vernonia colorata leaves with anti-inflammatory activity. AB - Bioassay-directed fractionation of an anti-inflammatory CHCl(3)-MeOH (9:1) extract of leaves of Vernonia colorata, using a carrageenan-induced rat paw model, led to the isolation of six new compounds (1-6). These were assigned as two new androst-8-en glycosides, 3-O-[beta-d-galactopyranosyl-(1-2)-[beta-d glucopyranosyl-(1-->6)]-beta-d-glucopyranoside]-5alpha,14alpha-androst-8-ene (1) and 3-O-[beta-d-glucopyranosyl-(1-->6)-beta-d-glucopyranoside]-5alpha,14alpha androst-8-ene (2), two new stigmastane-type glycosides, 3beta,21,24-trihydroxy 21,23;22,28;26,28-triepoxy-5alpha-stigmasta-8(9),14(15)-dien-3-O-beta-d galactopyranosyl-(1-->2)-beta-d-glucopyranoside (3) and 3beta,21,24-trihydroxy 21,23;22,28;26,28-triepoxy-5alpha-stigmasta-8(9),14(15)-dien-3-O-beta-d galactopyranosyl-(1-->2)-beta-d-(6-acetyl)glucopyranoside (4), and two new stigmastane-type steroids, 3beta,25,29-trihydroxy-5alpha-stigmasta 8(9),14(15),24Z(28)-triene (5) and 3beta,23,25-trihydroxy-24,28-epoxy-5alpha stigmasta-8(9),14(15)-diene (6). The structures of 1-6 were elucidated by spectral and chemical studies. Compounds 1-6 were tested for the anti inflammatory activity, but all were inactive or weakly inactive as anti inflammatory agents. PMID- 15043417 TI - Novel caprolactones from a marine streptomycete. AB - Two new caprolactones, (R)-10-methyl-6-undecanolide (1) and (6R,10S)-10-methyl-6 dodecanolide (2), were identified in the lipid extract of a marine streptomycete (isolate B6007). Their structures were proposed on the basis of GC-MS experiments and proved by synthesis. The absolute configuration of the compounds was established by comparison of the natural and synthetic stereoisomers using chiral gas chromatography. These caprolactones show a moderate phytotoxicity and a promising activity against cancer cells with concomitant low general cytotoxicity. PMID- 15043418 TI - New salicylic acid and isoflavone derivatives from Flemingia paniculata. AB - A new salicylic acid derivative, 2-carboxy-3-(2-hydroxypropanyl)phenol (1), and four new isoflavones, 5,7,4'-trihydroxy-8-(1,1-dimethylprop-2-enyl)isoflavone (2), 5,7,2',4'-tetrahydroxy-8-(1,1-dimethylprop-2-enyl)isoflavone (3), 5,2',4' trihydroxy-4' ',4' ',5' '(xi)-trimethyl-4' ',5' '-dihydrofurano-(7,6,2' ',3' ')isoflavone (4), and 5,2',4'-trihydroxy-7-(3-methylbut-2-enyloxy)isoflavone (5), were isolated from the stem bark of Flemingia paniculata. The structures of these compounds were established unambiguously by spectroscopic data interpretation. The biogenetic pathways to 1 and 2-4 have been postulated. PMID- 15043419 TI - Microbial transformation of isosteviol and inhibitory effects on Epstein-Barr virus activation of the transformation products. AB - Microbial transformation of isosteviol (2), a beyerane-type diterpenoid obtained from stevioside (1) by acid hydrolysis, yielded 7beta-hydroxyisosteviol (3), 11beta-hydroxyisosteviol (5), and 12beta-hydroxyisosteviol (6) by the fungus Aspergillus niger, 17-hydroxyisosteviol (7) by the fungus Glomerella cingulata, and 3 and 7-oxoisosteviol (4) by the fungus Mortierella elongate. The five metabolites, 3-7, along with 1 and 2 were evaluated for their inhibitory effects on Epstein-Barr virus early antigen (EBV-EA) activation induced by the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) in Raji cells as a primary screening test for inhibitors of tumor promoters. All the diterpenes tested showed potent inhibitory effects, with the five metabolites 3-7 exhibiting more potent effects. PMID- 15043420 TI - Dimeric flavonol glycoside and galloylated C-glucosylchromones from Kunzea ambigua. AB - A novel dimeric flavonol glycoside linked through a methylene group, kunzeagin A (1), and six new chromone C-glucosides, kunzeachromones A-F (2-7), were isolated along with seven known compounds from the leaf extract of Kunzea ambigua. The structures of these compounds were elucidated on the basis of spectroscopic analyses and chemical properties. Kunzeachromones A-F provided additional examples of galloylated C-glucosidic chromones occurring in the Myrtaceae. Kunzeagin A (1) and major constituents of this plant (6-C- and 8-C glucosylchromones and their monogallates) exhibited potent inhibitory effects on activation of Epstein-Barr virus early antigen induced by 12-O tetradecanoylphorbol 13-acetate in Raji cells. PMID- 15043421 TI - Effects of natural flavones and flavonols on the kinase activity of Cdk5. AB - A number of natural and synthetic flavonoids have been assessed previously with regard to their effects on the activity of cyclin-dependent kinases (Cdk1 and -2) related to the inhibition of cell cycle progression. On the other hand, the Cdk5/p35 system is of major importance in neuronal migration phenomena and brain development, and its deregulation is implicated in neurodegenerative diseases, particularly Alzheimer's. Here we show that some natural flavonoids inhibit the activity of the Cdk5/p35 system in the micromolar range, while others are practically inactive. Ring B-unsubstituted and highly methoxylated flavones were inactive or gave irreproducible results, and 6-methoxyapigenin and 6 methoxyluteolin were the most potent Cdk5 complex inhibitors within this series, while the common flavonols kaempferol and quercetin showed intermediate behavior. The reported crystal structure of the Cdk5 complex with its activator p25 was used for docking studies, which also led to the identification of the two 6 methoxyflavones, kaempferol and quercetin, as well as the untested 6-methoxy derivatives of kaempferol and quercetin and the corresponding 6-hydroxy analogues as compounds exhibiting a good fit to the active site of the enzyme. PMID- 15043422 TI - Resveratrol derivatives and their role as potassium channels modulators. AB - A series of stilbenoid analogues of resveratrol (trans-3,4',5-trihydroxystilbene) with a stilbenic or a bibenzylic skeleton have been prepared by partial synthesis from resveratrol and dihydroresveratrol. The synthesized compounds have been evaluated for their ability to modulate voltage-gated channels. PMID- 15043423 TI - Flavonoid glycosides from Adina racemosa and their inhibitory activities on eukaryotic protein synthesis. AB - From the dried leaves, flowers, and twigs of Adina racemosa, five new flavonoid glycosides, quercetin 3-O-alpha-l-rhamnopyranosyl(1-->6)-(3-O-trans-p-coumaroyl) beta-d-galactopyranoside (1), quercetin 3-O-alpha-l-rhamnopyranosyl(1-->6)-[(4-O trans-p-coumaroyl)-alpha-l-rhamnopyranosyl(1-->2)]-(4-O-trans-p-coumaroyl)-beta-d galactopyranoside (2), kaempferol 3-O-alpha-l-rhamnopyranosyl(1-->6)-[(4-O-trans p-coumaroyl)-alpha-l-rhamnopyranosyl(1--> 2)]-(4-O-trans-p-coumaroyl)-beta-d galactopyranoside (3), quercetin 3-O-alpha-l-rhamnopyranosyl(1-->6)-[(4-O-trans-p coumaroyl)-alpha-l-rhamnopyranosyl(1-->2)]-(3-O-trans-p-coumaroyl)-beta-d galactopyranoside (4), and quercetin 3-O-alpha-l-rhamnopyranosyl(1-->6)-[(4-O trans-caffeoyl)-alpha-l-rhamnopyranosyl(1-->2)]-(3-O-trans-p-coumaroyl)-beta-d galactopyranoside (5), and eight known compounds were isolated. The structures of the new compounds were determined by spectroscopic and chemical means. Their inhibitory activities on protein synthesis were assessed. The new glycosides were found to be inhibitors of eukayrotic, but not prokaryotic, protein synthesis. PMID- 15043424 TI - Sesquiterpene coumarins from Ferula fukanensis and nitric oxide production inhibitory effects. AB - Six new sesquiterpene coumarin derivatives, 2,3-dihydro-7-hydroxy-2R,3R-dimethyl 2-[4,8-dimethyl-3(E),7-nonadien-6-onyl]furo[3,2-c]coumarin (4), fukanefuromarin A (5), fukanefuromarin B (6), fukanefuromarin C (7), fukanefuromarin D (8), and fukanemarin A (9), were isolated from a 80% aqueous methanol extract of the roots of Ferula fukanensis. The structures were elucidated on the basis of spectral evidence, especially heteronuclear multiple-bond connectivity (HMBC), nuclear Overhauser exchange spectroscopy (NOESY), and high-resolution MS. An extract of F. fukanensis (FFE) and the sesquiterpene coumarin 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- 15043425 TI - Antityrosinase principles and constituents of the petals of Crocus sativus. AB - Three new monoterpenoids, crocusatin-J (1), -K (2), and -L (3), and a new naturally occurring acid, (3S),4-dihydroxybutyric acid (4), together with 31 known compounds were isolated and identified from the methanol extract of the petals of saffron (Crocus sativus). Their structures were established by spectroscopic methods. Among them, crocusatin-K (2), crocusatin-L (3), and 4 hydroxy-3,5,5-trimethylcyclohex-2-enone (8) showed significant antityrosinase activity, and protocatechuic acid (18), kaempferol (24), and kaempferol 7-O-beta d-glucopyranoside (27) were more effective in scavenging alpha,alpha-diphenyl beta-picrylhydrazyl (DPPH) radicals than alpha-tocopherol. In addition, the mechanism of tyrosinase inhibition by crocusatin-K (2) is also discussed. PMID- 15043426 TI - Cyclonellin, a new cyclic octapeptide from the marine sponge Axinella carteri. AB - Cyclonellin (1), a new cyclic octapeptide, was isolated from an aqueous extract of the marine sponge Axinella carteri. Its structure was elucidated by interpretation of NMR spectral data of the intact compound and N-terminal Edman sequencing of linear peptide fragments obtained by partial hydrolysis of 1. The absolute configurations of the constituent amino acids were determined by acid hydrolysis, derivitization with FDAA, and LC-MS analyses. PMID- 15043427 TI - Homogentisic acid derivatives from Miliusa balansae. AB - The new homogentisic acid derivatives miliusol (1b) and miliusolide (2) from Miliusa balansae were isolated and structurally determined by spectroscopic means. The relative configurations of the new 1b and its known acetate 1a were established. Furthermore, the symmetric ether bis(2-hydroxyphenyl)methyl ether 3, which was isolated for the first time from a natural source, the known flavonoids pachypodol and chrysosplenol C, and sodium benzoate were identified. PMID- 15043428 TI - Periconicins, two new fusicoccane diterpenes produced by an endophytic fungus Periconia sp. with antibacterial activity. AB - Two new fusicoccane diterpenes, named periconicins A (1) and B (2), with antibacterial activities have been isolated by bioassay-guided fractionation from an endophytic fungus Periconia sp., collected from small branches of Taxus cuspidata. The structures of the new compounds were determined by combined spectroscopic methods. PMID- 15043429 TI - New cytotoxic naphthopyrane derivatives from Adenaria floribunda. AB - Bioassay-guided fractionation of an EtOAc/MeOH extract of Adenaria floribunda young leaves using MCF-7, H-460, and SF-268 cancer cell lines yielded four new active compounds named adenaflorins A-D (1-4). Their chemical structures were determined by spectroscopic means. Adenaflorin A (1) was the most cytotoxic. PMID- 15043430 TI - Cytotoxic compounds from Mundulea chapelieri from the Madagascar Rainforest. AB - Bioassay-guided fractionation of methanolic extracts of Mundulea chapelieri resulted in the isolation of two new flavonoids, isomundulinol (1) and 3-deoxy-MS II (2), in addition to the eight known flavonoids 8-(3,3-dimethylallyl)-5,7 dimethoxyflavanone, MS-II, mundulinol, mundulone, munetone, rotenolone, rotenone, and tephrosin, and one known sesquiterpenoid, 8alpha-acetoxyelemol. The structures of the new flavonoids 1 and 2 were determined by 1D and 2D NMR experiments. All the isolated compounds were tested for cytotoxicity against the A2780 human ovarian cancer cell line; rotenolone and rotenone were the most potent compounds isolated, with IC(50) values of 0.5 and 0.7 microg/mL, respectively. PMID- 15043431 TI - Ketene acetal and spiroacetal constituents of the marine fungus Aigialus parvus BCC 5311. AB - Aigialone (1) and aigialospirol (2), two structurally unique compounds, were isolated from the mangrove fungus Aigialus parvus BCC 5311. The structure of the new ketene acetal 1 was elucidated by spectral analysis, and its relative stereochemistry was determined by X-ray crystallography. The stereochemistry of aigialospirol (2), elucidated by NMR spectral analysis, suggested that this compound is possibly derived from hypothemycin (3), a metabolite previously isolated from this same fungus. PMID- 15043432 TI - Jusbetonin, the first indolo[3,2-b]quinoline alkaloid glycoside, from Justicia betonica. AB - A new indolo[3,2-b]quinoline alkaloid glycoside, jusbetonin (1), and three known alkaloids, namely, 10H-quindoline (2), 6H-quinindoline (3), and 5H,6H-quinindolin 11-one (4), have been isolated from the leaves of Justicia betonica. The structure of 1 was established on the basis of 1D and 2D NMR ((1)H-(1)H COSY, HMQC, and HMBC) and HRFABMS data. Compound 1 is the first example of a glycosylated indolo[3,2-b]quinoline alkaloid, while compound 4 was isolated for the first time from a natural source. PMID- 15043433 TI - Discorhabdins I and L, cytotoxic alkaloids from the sponge Latrunculia brevis. AB - Two cytotoxic alkaloids, discorhabdins L (1) and I (2), were isolated from Latrunculia brevis and their structures assigned on the basis of detailed spectroscopic analysis and comparison with the known metabolites discorhabdins R (5), D (6), and B (4). Compounds 1 and 2 showed strong in vitro cytotoxicity against a panel of 14 tumor cell lines. PMID- 15043434 TI - New peptaibols from Mycogone cervina. AB - From cultures of Mycogone cervina, a mycophilic fungus growing on the fruit bodies of ascomycete Paxina acetabulum, two new peptaibols (cervinin, N-Ac-Leu Aib-Pro-Aib-Leu-Aib-Pro-Ala-Aib-Pro-Val-red-Leu (1) and the red-Leu O-acetylated derivative 2) were isolated. The structures of 1 and 2 were elucidated by spectroscopic techniques. They exhibited weak antibacterial as well as cytotoxic activities. They are the first secondary metabolites described from M. cervina. PMID- 15043435 TI - Four new triterpenoids from Maytenus ilicifolia. AB - Four new triterpenoids with various skeletons, maytefolins A-C (1-3) and uvaol-3 caffeate (4), were isolated from the leaves of a Brazilian medicinal plant, Maytenus ilicifolia, together with five known triterpenoids. Of these triterpenoids only erythrodiol exhibited significant cytotoxicity against KB/S, KB/VJ300, and KU 19-20 cells. PMID- 15043436 TI - Isolation and characterization of polybrominated diphenyl ethers as inhibitors of microtubule assembly from the marine sponge Phyllospongia dendyi collected at Palau. AB - Two new polybrominated diphenyl ethers (1 and 2) were isolated by bioassay-guided separations together with nine known compounds (3-11) from the marine sponge Phyllospongia dendyi collected from Palau. The structures were assigned on the basis of their spectral data. Compounds 3, 7, and 10 showed inhibitory activities to the assembly of microtubule proteins (IC(50): 29.6, 33.5, and 20.9 microM, respectively) and to the meiotic maturation of starfish oocytes (IC(50): 3.6, 4.2, and 4.2 microM, respectively), while 1, 2, 4-6, 8, 9, and 11 were not active at 100 microM. Two phenolic hydroxyls are required for their bioactivities. PMID- 15043438 TI - (+)- and (-)-syn-2-isobutyl-4-methylazetidine-2,4-dicarboxylic acids from the extract of Monascus pilosus-fermented rice (red-mold rice). AB - The structures of two enantiomeric azetidine-type amino acids isolated from the n butanol-soluble fraction of the 70% ethanol extract of red-mold rice fermented with Monascus pilosus were established to be (+)- [1; (+)-monascumic acid] and ( )-syn-2-isobutyl-4-methylazetidine-2,4-dicarboxylic acids [2; (-)-monascumic acid] based on spectroscopic methods. PMID- 15043437 TI - Scleritodermin A, a cytotoxic cyclic peptide from the lithistid sponge Scleritoderma nodosum. AB - The lithistid sponge Scleritoderma nodosum contains a new cyclic peptide, scleritodermin A (1), the structure of which incorporates l-proline, l-serine, and keto-allo-isoleucine units as well as a novel conjugated thiazole moiety (ACT) and O-methyl-N-sulfoserine. Scleritodermin A (1) inhibited tubulin polymerization and showed significant in vitro cytotoxicity against human tumor cell lines. PMID- 15043439 TI - Phenolic metabolites of Dalea versicolor that enhance antibiotic activity against model pathogenic bacteria. AB - A new flavonoid (1) was isolated from organic extracts of Dalea versicolor, along with six known phenolic compounds (2-7). The structures of the seven compounds were determined by NMR and HRMS methods. These compounds were evaluated for direct activity against a variety of organisms in vitro, including the Gram positive bacteria Staphylococcus aureus and Bacillus cereus. In addition, the compounds were evaluated for their ability to potentiate the activity of known antimicrobials through inhibition of multidrug-resistance (MDR) pumps. Compounds 1, 2, 4, and 7 exhibited direct or synergistic activity toward the human pathogen S. aureus and the opportunistic pathogen B. cereus. Compounds 4 and 7 were also found to potentiate the activity of berberine and of prescribed antibiotics, with 4 demonstrating a mode of action consistent with inhibition of the NorA MDR efflux pump in S. aureus. PMID- 15043440 TI - Lakoochins A and B, new antimycobacterial stilbene derivatives from Artocarpus lakoocha. AB - Two new stilbene derivatives, lakoochins A (1) and B (2), were isolated from the roots of Artocarpus lakoocha. The structures of 1 and 2 were elucidated by analysis of their spectral data. Lakoochins A (1) and B (2) exhibited antimycobacterial activity with the respective MIC values of 12.5 and 50 microg/mL. While 1 was cytotoxic against the BC (breast cancer) cell line (IC(50) 6.1 microg/mL) but inactive (at 20 microg/mL) toward KB (nasopharyngeal carcinoma) cells, compound 2 possessed cytotoxicity against the BC and KB cell lines with IC(50) values of 3.1 and 6.1 microg/mL, respectively. PMID- 15043441 TI - Acetals of three new cycloartane-type saponins from Egyptian collections of Astragalus tomentosus. AB - Three new cycloartane-type saponin ethyl acetals, deacetyltomentoside I (2), tomentoside III (3), and tomentoside IV (4), were isolated along with the known acetal tomentoside I (1) from the aerial parts of Astragalus tomentosus of Egyptian origin. The saponins from which the acetals are most probably derived are also new compounds. The structures of the acetals were established as 6alpha hydroxy-23alpha-ethoxy-16beta,23(R)-epoxy-24,25,26,27-tetranor-9,19-cyclolanosta 3-O-beta-d-xyloside (2), 6alpha-acetoxy-23alpha-ethoxy-16beta,23(R)-epoxy 24,25,26,27-tetranor-9,19-cyclolanosta-3-O-[beta-d-(4'-trans-2-butenoyl)]xyloside (3), and 6alpha-acetoxy-23alpha-ethoxy-16beta,23(R)-epoxy-24,25,26,27-tetranor 9,19-cyclolanosta-3-O-[beta-d-glucopyranosyl(1 --> 2)]-beta-d-xyloside (4), by detailed spectroscopic and chemical studies. PMID- 15043442 TI - New cytotoxic steroids from the Indian Ocean sponge Axinella cf. bidderi. AB - Four new sterols have been isolated from the marine sponge Axinella cf. bidderi, 17alpha-hydroxy-22,23-epoxy-24-methylcholest-5-en-3beta-ol (1) and 17alpha hydroxy-22,23-epoxycholest-5-en-3beta-ol (2), together with 3 and 4, which possess respectively the cholestene and the cholestane skeleton with a cyclic enol ether linkage between C-18 and C-22. The structures were elucidated using spectroscopic data. The in vitro activity was evaluated against prostate, ovary, pancreas, colon, and lung cell lines. PMID- 15043443 TI - Novel meroditerpenes from the brown alga Cystoseira sp. AB - Five new meroditerpenes have been isolated from a brown alga of the genus Cystoseira collected around the Canary Islands. One, cystoseirone diacetate (3), possesses a new rearranged structure with an unusual ether linkage in the diterpene side chain. Its biogenetic origin was explained as derived from the oxidation of amentol chromane diacetate (2) and subsequent cyclization. Structures were determined through the interpretation of the spectral data and by means of chemical transformations. The relative stereochemistry was proposed on the basis of ROESY correlations. PMID- 15043444 TI - Steroidal alkaloid glycosides from tomato (Lycopersicon esculentum). AB - Three new steroidal alkaloid glycosides, lycoperosides F-H (1-3), were isolated from tomato fruits (Lycopersicon sculentum) along with lycoperosides A-D, esculeoside A, and rutin. The structures of these glycosides were characterized as the 3-O-beta-lycotetraosides of 23(R)-23-acetoxy-27-hydroxy-27-O-beta-d glucopyranosyltomatidine (1), (23S,24R)-23-acetoxy-24-O-beta-d glucopyranosylsoladulcidine-24-ol (2), and 22-isopimpifolidine (3), by means of their spectroscopic data. Also obtained was the new natural product lycoperodine 1 (4). PMID- 15043445 TI - New nitrogenous Germacranes from a Thai marine sponge, Axinyssa n. sp. AB - Two new germacrane sesquiterpenes, including (1Z,4Z)-7alphaH-11-aminogermacra 1(10),4-diene (1) and N,N'-11-bis[(1Z,4Z)-7alphaH-germacra-1(10),4-dienyl]urea (2), were isolated from a Thai marine sponge Axinyssan. sp. collected from the Andaman Sea. The structure elucidations of 1 and 2 were accomplished by means of 1D and 2D NMR, MS, and IR spectroscopy. Only 1 exhibited strong antimicrobial activity against Staphylococcus aureus, Bacillus subtilis, and Candida albicans. PMID- 15043446 TI - Antineoplastic agents. 380. Isolation and X-ray crystal structure determination of isoaaptamine from the Republic of Singapore Hymeniacidon sp. and conversion to the phosphate prodrug hystatin 1. AB - By use of bioassay (murine P388 lymphocytic leukemia cell line) guided isolation procedures, extracts of the Republic of Singapore marine sponge Hymeniacidon sp. were found to contain demethyloxyaaptamine (1) and aaptamine (3) as prominent cancer cell growth inhibitory constituents accompanied by the trace, albeit more active, component isoaaptamine (4). The isolation, X-ray structure elucidation, and antineoplastic and antimicrobial activities of isoaaptamine (4) have been summarized. Because of instability, isoaaptamine (4) was converted to a stable sodium phosphate prodrug designated hystatin 1 (7). PMID- 15043447 TI - Challenges and rewards of research in marine natural products chemistry in Brazil. AB - Brazil is blessed with a great biodiversity, which constitutes one of the most important sources of biologically active compounds, even if it has been largely underexplored. As is the case of the Amazon and Atlantic rainforests, the Brazilian marine fauna remains practically unexplored in the search for new biologically active natural products. Considering that marine organisms have been shown to be one of the most promising sources of new bioactive compounds for the treatment of different human diseases, the 8000 km of the Brazilian coastline represents a great potential for finding new pharmacologically active secondary metabolites. This review presents the status of marine natural products chemistry in Brazil, including results reported by different research groups with emphasis on the isolation, structure elucidation, and evaluation of biological activities of natural products isolated from sponges, ascidians, octocorals, and Opistobranch mollusks. A brief overview of the first Brazilian program on the isolation of marine bacteria and fungi, directed toward the production of biologically active compounds, is also discussed. The current multidisciplinary collaborative program under development at the Universidade de Sao Paulo proposes to establish a new paradigm toward the management of the Brazilian marine biodiversity, integrating research on the species diversity, ecology, taxonomy, and biogeography of marine invertebrates and microorganisms. This program also includes a broad screening program of Brazilian marine bioresources, to search for active compounds that may be of interest for the development of new drug leads. PMID- 15043450 TI - Cytologic features of squamous cell carcinoma in ThinPrep slides: evaluation of cases that performed poorly versus those that performed well in the College of American Pathologists Interlaboratory Comparison Program in Cervicovaginal Cytology. AB - CONTEXT: Although the cytologic features of squamous cell carcinoma in ThinPrep specimens are well known, whether these features are different in cases that are easily identified than in cases that are more difficult to identify is not known. OBJECTIVE: To determine the cytologic features of squamous cell carcinoma in ThinPrep specimens that are easy to identify versus those that are difficult. DESIGN: The cytologic features of 6 cases of squamous cell carcinoma that performed poorly in the College of American Pathologists Interlaboratory Comparison Program were compared with 14 cases that performed extremely well. RESULTS: After evaluation of multiple criteria, 7 different cytologic features were analyzed based on review by a consensus panel blinded to the performance of the cases. The feature that was most strongly associated with cases that performed poorly was the presence of Trichomonas vaginalis (5/6 [83%] vs 0/14; P <.001). The presence of marked nuclear pleomorphism was more common in cases that performed well (4/14 [28%] vs 0/6; P =.27), but was not significant. The number of tumor cells, the number of normal cells, and the presence of keratinization, pleomorphism, nucleoli, and diathesis were not significant. The most common misdiagnosis after Trichomonas vaginalis was reparative change. CONCLUSIONS: The presence of Trichomonas is characteristic of cases of squamous cell carcinoma in ThinPrep slides that are often misdiagnosed in this program. While Trichomonas is identified by participants in some of these cases, a significant percentage of participants interpreted the findings as reparative, without identifying the organism. These results emphasize the importance of distracting factors, whether identified or not, in evaluating gynecologic cytology. PMID- 15043451 TI - Heparin monitoring and patient safety: a College of American Pathologists Q Probes study of 3431 patients at 140 institutions. AB - CONTEXT: Appropriate laboratory monitoring of unfractionated heparin therapy promotes effective anticoagulation while minimizing hemorrhagic complications. OBJECTIVES: To measure heparin therapy monitoring in a "real-world" setting and to assess the degree of anticoagulation achieved. DESIGN: One hundred forty institutions abstracted laboratory and pharmacy data from up to 30 inpatients receiving standard-dose unfractionated heparin therapy for 72 hours. Institutions also reported their therapeutic ranges and described heparin prescribing and monitoring policies. RESULTS: Activated partial thromboplastin times or anti factor Xa levels were measured at least once within the first 12 hours of administration for 95% of 3431 heparinized inpatients. Eighty-seven percent of patients had a platelet count performed within 72 hours of heparin administration. Seventy-eight percent of heparinized inpatients achieved therapeutic anticoagulation within 24 hours, but more than one third of patients entered the supratherapeutic range on at least 2 occasions during the first 72 hours. We found moderate variation in performance among the 140 institutions participating in the study, with more consistency in monitoring patients but less consistency in achieving therapeutic levels of anticoagulation. In one fourth of hospitals, more than half of the heparinized patients entered the supratherapeutic range on 2 or more occasions during the first 72 hours of therapy. None of 20 institutional practices we examined were meaningfully associated with more thorough monitoring of patients or with a higher percentage of patients achieving therapeutic anticoagulation. There was moderately wide variation in therapeutic ranges among the 140 sites. CONCLUSIONS: The prevention of heparin over-anticoagulation represents an important opportunity for improving patient safety in a significant number of institutions. PMID- 15043454 TI - Giant cell glioblastoma and pleomorphic xanthoastrocytoma. PMID- 15043456 TI - Three cases of massive fetomaternal hemorrhage presenting without clinical suspicion. AB - Fetomaternal hemorrhage (FMH) is a common obstetrical occurrence most often associated with small volumes of blood transferred across the placenta. Fetomaternal hemorrhage leads to alloimmunization of Rh D-negative mothers, resulting in an increased risk of hemolytic disease of the newborn. Massive FMH involving volumes of blood greater than 30 mL can cause substantial fetal morbidity and mortality. Massive FMH may present with signs and symptoms such as decreased movement, sinusoidal heart rhythms, or fetal anomalies. We present 3 cases of clinically unexpected massive FMH of 206, 88, and 155 mL. The treating clinicians were unaware of any fetal or maternal signs or symptoms of FMH until contacted by the laboratory. These cases illustrate the necessity for FMH quantitation, even in the absence of clinical suspicion. Additional studies are needed to find better ways to identify these patients in advance. Development of criteria allowing identification of patients at risk would be of benefit to both mother and baby. PMID- 15043457 TI - Recurrent solitary fibrous tumor of the pleura with malignant transformation. AB - Recurrent solitary fibrous tumor of the pleura with malignant progression occurs rarely. We report a case of solitary fibrous tumor of the pleura in an 85-year old white woman that recurred 4 times during a span of 10 years and subsequently underwent malignant transformation. The accurate diagnosis of solitary fibrous tumor is aided by ancillary techniques, such as immunohistochemical staining; however, with malignant transformation, such tools may be of limited value. Long term clinical follow-up is recommended for all patients with solitary fibrous tumor because of the potential adverse biological behavior of this tumor, which may lead to repeated recurrences and/or malignant transformation. PMID- 15043458 TI - Urachal malignant fibrous histiocytoma: a case report and review of the literature. AB - Pathologic processes involving the urachus are usually related to inflammatory or sinofistular conditions. Neoplasms rarely arise within this structure, and when they do occur, they are typically epithelial, with mucinous adenocarcinoma being the most common. Mesenchymal lesions, both benign and malignant, have rarely been described in this location. We report the case of a 66-year-old white man who presented with a primary urachal malignant fibrous histiocytoma and died of metastatic disease 20 months after the initial diagnosis. This is an unusual case of malignant fibrous histiocytoma arising in a urachal remnant. PMID- 15043459 TI - Giant cell tumor of rib masquerading as thymoma: a diagnostic pitfall in needle core biopsy of the mediastinum. AB - Giant cell tumor of bone is rarely seen in the rib, where it may present as a mediastinal mass. The diagnosis of giant cell tumor of bone is generally straightforward by fine-needle aspiration or needle core biopsy, but sampling problems may lead to confusion with other neoplasms or inflammatory processes. Here, we report a case of giant cell tumor of rib presenting as a mediastinal mass in a 36-year-old man. Because of inadequate sampling and inaccurate clinical information, the tumor was initially mistaken for thymoma. When the mass failed to respond to conventional chemotherapy, additional tissue was obtained and a giant cell tumor was diagnosed. Consequently, definitive therapy was delayed. The case illustrates an important diagnostic pitfall in the biopsy of mediastinal masses. PMID- 15043460 TI - Biphasic acute myeloid leukemia with near-tetraploidy and immunophenotypic transformation. AB - This report describes a case of acute myeloid leukemia (subtype M1) with biphasic morphology. The bone marrow biopsy showed 2 distinct regions of blasts, one containing large cells and the other small cells. Morphometric and DNA ploidy analysis showed that the mean nuclear area and mean DNA index for the large cell region were 2-fold higher than those for the small cell region. Cytogenetic analysis showed an abnormal near-tetraploid clone. The tumor relapsed following aggressive therapy. The cells from the relapse specimen were similar to the original small cell region with respect to nuclear area and DNA index; however, there was immunophenotypic transformation with gain of CD7 and gain of CD56. Cytogenetically, the relapse specimen showed no evidence of the near-tetraploid clone, but instead had a previously unidentified abnormal clone containing 46 chromosomes and structural abnormalities of 2q and 7q. Biphasic morphology in acute myeloid leukemia may be predictive of a near-tetraploid subclone and immunophenotypic transformation. PMID- 15043461 TI - Inlet patch: prevalence, histologic type, and association with esophagitis, Barrett esophagus, and antritis. AB - CONTEXT: Inlet patch is a congenital anomaly of the cervical esophagus consisting of gastric mucosa. Case reports have documented the histologic type and its associated complications. OBJECTIVE: To report the prevalence and histologic types of inlet patch as well as its association with Barrett esophagus and Helicobacter pylori-associated gastritis. DESIGN: We reviewed 1821 consecutive pathology reports from endoscopies of the upper gastrointestinal tract between 1995 and 2002 and identified 20 patients with inlet patch. The patients' ages ranged from 16 to 75 years (mean, 55 years). We examined biopsies from these patients of the cervical esophagus, distal esophagus, and antrum that had been stained with hematoxylin-eosin and the Steiner stain. Biopsies from the cervical esophagus composed of nonoxyntic gastric mucosa were also stained for gastrin producing cells. RESULTS: In our patient population, inlet patch occurred in 1.1% of all patients in whom an inlet patch had been searched for endoscopically and a biopsy performed. In the inlet patch, oxyntic mucosa was the most common histologic type (11/20), followed by cardiac mucosa (5/20). Four specimens of the inlet patch contained only foveolar epithelium and were therefore considered too superficial to be classified. Twelve of 20 biopsies of the inlet patch were inflamed, and 1 of them was associated with H pylori. Pancreatic acinar tissue was noted in 2 patients; no intestinal metaplasia was found. In the distal esophagus, 4 patients with inlet patch had distal esophagitis, 4 had Barrett esophagus, 5 had oxyntic mucosa, 3 had pancreatic acinar tissue (1 coexisting with oxyntic mucosa), and 5 presented with only unremarkable esophageal squamous mucosa. Antral gastritis was seen in 2 patients, 1 of them with H pylori. The same patient also had H pylori in the inlet patch. CONCLUSION: Inlet patch occurred in about 1% of our study population. Oxyntic mucosa constituted the most common histologic type; inflammation was common. The H pylori infection of the inlet patch correlated with that of the antrum. None of the inlet patch biopsies showed intestinal metaplasia. Pancreatic acinar tissue occurred with similar frequency in the inlet patch and distal esophagus. Esophagitis was noted in 25% of the patients with inlet patch, and Barrett esophagus was noted in 20%. PMID- 15043462 TI - Malignant gastrointestinal leiomyosarcoma and gastrointestinal stromal tumor with prominent osteoclast-like giant cells. AB - CONTEXT: One case of leiomyosarcoma and one case of gastrointestinal stromal tumor with prominent osteoclast-like giant cells have so far been reported in the digestive tract. OBJECTIVE: To ascertain the clinicopathologic features and biologic behavior of these tumors, we report 3 additional cases of leiomyosarcoma of the gastrointestinal tract and one malignant gastrointestinal stromal tumor. DESIGN: Histologic and immunohistochemical examinations were performed. Clinical and follow-up data were recorded, and the literature was reviewed. RESULTS: The age of the patients ranged from 50 to 68 years (mean, 62 years). One of the lesions arose in the stomach, one in the ileum, and 2 in the colon. Three tumors showed a strong positivity for muscle actin and desmin and were diagnosed as leiomyosarcomas, 2 of them showing spindle cells and 1 of them showing epithelioid cells. The fourth tumor reacted strongly positive for c-Kit (CD117) and vimentin, and it was diagnosed as an epithelioid malignant gastrointestinal stromal tumor. All tumors were characterized by numerous osteoclast-like giant cells that were unevenly distributed and that, using immunohistochemistry, reacted strongly with CD68. CONCLUSIONS: Malignant stromal tumors with osteoclast like giant cells of the gastrointestinal tract are rare entities, are more commonly of a myogenic origin such as leiomyosarcoma, and seem to have an aggressive behavior. PMID- 15043463 TI - Hemoglobin C and hemoglobin O-Arab variants can be diagnosed using the Bio-Rad Variant II high-performance liquid chromatography system without further confirmatory tests. AB - CONTEXT: Current standards for laboratory accreditation from the College of American Pathologists state that when high-performance liquid chromatography (HPLC) is used as a screening test, all non-A, non-S abnormal hemoglobin (Hb) variants must be confirmed by an alternative method, including alkaline and acid electrophoresis. OBJECTIVE: To determine whether confirmation of Hb C and Hb O Arab variants by an alternative method is required when using the Bio-Rad Variant II HPLC system. DESIGN: We reviewed 48 478 consecutive hemoglobin identification test results performed on the Bio-Rad Variant II HPLC system during the period November 15, 2000 to January 15, 2003. SETTING: Special Hematology Laboratory, Department of Pathology, Bellevue Hospital Center, New York, NY. MAIN OUTCOME MEASURES: The chromatogram patterns and retention times (RTs) for specimens containing Hb C and Hb O-Arab were analyzed. We compared the results by the HPLC method with those by the confirmatory tests (alkaline and acid electrophoresis) for both variants. RESULTS: We identified 3668 cases of abnormal hemoglobin variants, including 660 cases of Hb C trait (17%), 5 cases of Hb O-Arab trait (0.1%), and 1 case of Hb SO-Arab (0.03%). A unique pattern of separation on the chromatogram for Hb O-Arab was revealed, presenting as 2 distinct peaks in 2 different manufacturer-defined RT windows, namely, D and C. The chromatogram for Hb C did not show the D window in any of the reviewed cases. The RT in the C window (C-RT) revealed a statistically significant difference for Hb C and Hb O Arab (5.18 +/- 0.01 minutes and 4.91 +/- 0.01 minutes, respectively; P <.001). CONCLUSION: According to our review, the identification of Hb C and Hb O-Arab is accurate using HPLC methodology, as performed by the Bio-Rad Variant II HPLC system. This method can be both confirmatory and diagnostic at the same time. PMID- 15043464 TI - Diagnostic and prognostic value of cardiac troponin I assays in patients admitted with symptoms suggestive of acute coronary syndrome. AB - CONTEXT: Increasing numbers of patients are presenting to emergency departments with symptoms suggestive of an acute myocardial infarction. OBJECTIVE: To demonstrate the comparative performance of the Ortho Vitros Troponin I and Beckman Access AccuTnI assays used to detect myocardial infarction and to develop risk stratification schemes for all-cause death in patients who presented with myocardial ischemia symptoms that were suggestive of acute coronary syndrome (ACS). DESIGN: The prospective enrollment of patients with ACS and the measurement of serial plasma samples by 2 commercial cardiac troponin I (cTnI) assays. SETTING: A metropolitan medical center that admitted patients with ACS during a 2-month period. PATIENTS: The study population consisted of 200 consecutively admitted patients who presented with symptoms that were suggestive of ACS. RESULTS: Correlation scatterplots showed no significant bias between cTnI assays based on 659 specimens across the dynamic range of each assay. Only minor differences in slopes and intercepts were observed between assays when correlations were based across selected concentration ranges. The receiver operating characteristic curve areas for the detection of myocardial infarction were not significantly different (Ortho,.991; Beckman,.995). At the 99th percentile (Beckman, 0.04 microg/L; Ortho, 0.08 microg/L), each assay demonstrated 100% sensitivity with 78% and 80% specificity, respectively. Kaplan Meier survival curves and the log-rank test were used to compare time-to-event data. Patients with increased baseline cTnI values had higher odds ratios of death than did those with normal concentrations. For Ortho, the 99th percentile cutoff was 5.9, and the 10% coefficient of variation cutoff was 10.3; for Beckman, the 99th percentile cutoff was 31.4, and the 10% coefficient of variation cutoff was 15.3. CONCLUSIONS: Comparable diagnostic and risk stratification abilities were demonstrated in patients with ACS by the Ortho Vitros and Beckman Access cTnI assays, with no significant analytic bias between cTnI assays. PMID- 15043465 TI - Cellular proliferative fraction measured with topoisomerase IIalpha predicts malignancy in endocrine pancreatic tumors. AB - CONTEXT: Endocrine pancreatic tumors (EPTs) are rare lesions with varying biological behavior. Establishing malignancy is a challenge for clinicians and pathologists. OBJECTIVE: To establish the role of proliferative, apoptotic, angiogenic, and hormonal markers as predictors of malignancy in EPTs. DESIGN: Paraffin-embedded EPT samples were studied for prognostic markers. PATIENTS: Twenty-one consecutive patients with a diagnosis of EPT. MAIN OUTCOME MEASURES: The proliferative fraction (topoisomerase IIalpha), microvascular density (CD34), vascular endothelial growth factor expression, and estrogen receptor-beta (ERbeta) expression were studied by immunohistochemistry on all EPTs. Apoptosis was also assessed with terminal deoxynucleotidyl transferase nick-end labeling. RESULTS: We identified 13 benign and 8 malignant tumors. Topoisomerase IIalpha was significantly increased in malignant tumors (P =.001), while there were no differences in apoptosis, microvascular density, or vascular endothelial growth factor expression in association with malignancy. No correlation could be identified between microvascular density and vascular endothelial growth factor expression, and ERbeta was not detected. A receiver operating characteristic curve for topoisomerase IIalpha disclosed that above a labeling index of 13, the test had 88% sensitivity and 100% specificity for predicting malignancy. CONCLUSION: Cellular proliferation measured with topoisomerase IIalpha is a simple prognostic marker for malignancy in EPTs, unlike apoptosis, angiogenesis, or the presence of ERbeta, which were not associated with malignant behavior. These findings designate a defined field for future research on endocrine pancreatic carcinogenesis and a possible target for chemotherapeutic agents. PMID- 15043466 TI - Role of PTEN in gastrointestinal stromal tumor progression. AB - CONTEXT: Gastrointestinal stromal tumors (GISTs) are Kit/CD117-expressing mesenchymal neoplasms of uncertain malignant potential. The lack of a reliable method of prognostication hampers the selection of patients eligible for STI571 therapy. 10q22-q23 is a region involved in chromosomal losses found in a fraction of malignant primary and metastatic GISTs harboring PTEN (phosphatase and tensin homologue deleted on chromosome 10), a tumor suppressor gene often altered in human neoplasms. OBJECTIVE: To investigate the role of PTEN in GISTs, an issue that to our knowledge has not been addressed previously. DESIGN: PTEN status was determined in a series of 21 GISTs, with follow-up ranging between 6 and 198 months, using immunohistochemistry correlated with clinical data. RESULTS: A greater than 25% fraction of cells with low or absent PTEN immunostaining was detected in 9 GISTs, including all those showing malignancy. By the log-rank test, a fraction of PTEN-deficient cells greater than 25% was associated with malignancy (P <.001). Percentage of cells underexpressing PTEN, size, cellularity, MIB-1 immunoreactivity, and coagulative necrosis proved to be associated with malignancy by Cox proportional hazards univariate analysis; low or absent expression of PTEN was the only factor selected by multivariate analysis (P =.03). CONCLUSIONS: PTEN downregulation is implied in GIST progression. The immunohistochemical assessment of PTEN status appears to be a promising method of GIST prognostication. PMID- 15043467 TI - Evaluation of a dimeric inhibin-A assay for assessing fetal Down syndrome: establishment, comparison, and monitoring of median concentrations for normal pregnancies. AB - CONTEXT: Several studies report the role of dimeric inhibin-A in assessing risk for fetal Down syndrome. The majority, however, use the Serotec inhibin-A assay and not the newer Diagnostic Systems Laboratories inhibin-A enzyme-linked immunosorbent assay (ELISA). OBJECTIVES: To establish normal gestational age day specific medians, to compare our results against previous studies pertaining to the inhibin-A ELISA, and to evaluate long-term assay performance. DESIGN: Using the inhibin-A ELISA, 100 specimens were assayed for each completed week of gestation for weeks 15 to 20, 50 specimens for 14 weeks, and 54 specimens for 21 weeks or older. Regressed inhibin-A medians were calculated employing a second degree polynomial fit of the arithmetic medians. Thereafter, inhibin-A ELISA lot comparisons were performed to evaluate consistency. RESULTS: Regressed values of 182, 174, 175, 184, 201, and 226 pg/mL resulted for weeks 15 to 20, respectively [pg/mL inhibin-A = 4.1528(gestational age)2 - 136.49(gestational age) + 1294.9]. A comparison with 2 other studies shows our values to be lower overall by 15 +/- 11.4% and 16 +/- 2.6%. However, variability between kit lots was as high as 30%. CONCLUSIONS: The equation derived provides for the calculation of gestational age day-specific inhibin-A medians for integration into maternal serum screening programs with a subsequent decrease in false-positives expected and observed. Our medians differ considerably from those of other studies, with limited data, using the Diagnostic Systems assay. However, lot changes since the initial analysis have exhibited similar inconsistencies. Therefore, we recommend that others incorporating the assay into their screening programs carefully establish, monitor, and adjust their medians accordingly as a result of potential variations. PMID- 15043469 TI - Pathologic quiz case: multiple congenital birth defects in a full-term infant. Urorectal septum malformation sequence. PMID- 15043468 TI - Immunohistochemical analysis of lung carcinomas with pure or partial bronchioloalveolar differentiation. AB - CONTEXT: In 1999, the World Health Organization redefined bronchioloalveolar carcinomas (BACs) as those neoplasms with only a pure lepidic growth pattern and no invasion. OBJECTIVES: The present study examined 45 lung cancers with a BAC component (1) to determine whether these tumors would be classified as BACs by current World Health Organization standards, (2) to quantitate the BAC component within these tumors, and (3) to see if phenotypic differences exist between the so-called invasive and noninvasive regions of these tumors. DESIGN: Retrospective review of hematoxylin-eosin-stained slides and classification of histologic grade, tumor subtype, and percentage of pure BAC pattern, with further characterization by immunohistochemical staining for thyroid transcription factor 1, cytokeratin 7, cytokeratin 20, and Ki-67 antibodies. RESULTS: Only 7 (15.6%) of the 45 tumors examined could be classified as BAC by current strict World Health Organization criteria. Those tumors, classified as nonmucinous and mixed, showed similar immunohistochemical staining for cytokeratin 7, cytokeratin 20, and thyroid transcription factor 1; mucinous tumors showed disparate staining. Significant differences in immunohistochemical staining and tumor cell proliferation were seen for the regions of tumors designated as lepidic, infiltrative, and leading edge and for the regions of tumors with different histologic grades (ie, well, moderately, and poorly differentiated). CONCLUSIONS: Nonmucinous and mixed BACs are phenotypically similar and show identical immunohistochemical staining patterns; mucinous tumors, on the other hand, show disparate immunohistochemical staining. Pulmonary neoplasms designated as adenocarcinomas with a BAC component represent a heterogenous group with a range of cell types, differentiation, growth, and immunophenotypes. Within an individual neoplasm, there are regional differences in these parameters as well. PMID- 15043470 TI - Pathologic quiz case: a 54-year-old deceased man with diffuse subcortical lesions of the central nervous system. Progressive multifocal leukoencephalopathy. PMID- 15043471 TI - Pathologic quiz case: a 77-year-old woman with bilateral breast masses. Amyloidosis involving the breast. PMID- 15043472 TI - Pathologic quiz case: myxoid tibial lesion in a 31-year-old man. Low-grade myxofibrosarcoma. PMID- 15043473 TI - Pathologic quiz case: a 29-year-old woman with thrombocytosis. Chronic myelogenous leukemia. PMID- 15043474 TI - Prostate gland-like epithelium in the epididymis: a case report and review of the literature. AB - Aberrant prostatic tissue occurs commonly in the adult male urethra and bladder. Ectopic prostatic tissue occurring outside the urinary system is rare. One case with scattered prostate-type glands in epididymis has been reported in the literature. We report a related case, in which the presence of prostate gland like epithelium was recognized in epididymal glands in routine histology and was confirmed by subsequent immunohistochemical analysis using prostate-specific antigen. We then examined 23 additional orchiectomy specimens for the presence of prostate-like epithelium. The possibility of this being true ectopia versus an unusual metaplastic change is discussed. PMID- 15043475 TI - Primary adenocarcinoma of cutaneous vesicostomy 40 years later: a rare case. AB - We present a case of adenocarcinoma developing at the vesicocutaneous edge of a vesicostomy, 40 years after it was created, in a patient who underwent cadaveric kidney transplant. Although transitional and squamous cell carcinoma of a vesicostomy have been reported, to our knowledge, the presence of adenocarcinoma at the vesicostomy edge has not been reported previously. PMID- 15043477 TI - Coexistent hereditary hemochromatosis and alpha1-antitrypsin deficiency. PMID- 15043478 TI - Importance of immunohistochemical staining in metastatic anaplastic oligodendroglioma. PMID- 15043479 TI - Pathologic quiz case: a 49-year-old man with left parotid mass. Adenocarcinoma, not otherwise specified. PMID- 15043480 TI - Pathologic quiz case: an asymptomatic 79-year-old woman with an abnormal Papanicolaou test. Adenoid basal carcinoma of the cervix. PMID- 15043481 TI - Pathologic quiz case: a 27-year-old man with abdominal pain. Peutz-Jeghers syndrome. PMID- 15043482 TI - Pathologic quiz case: a slow and awkward child. Central core disease. PMID- 15043483 TI - Musculoskeletal conditions affect millions. PMID- 15043484 TI - Analysis of on-line clinical laboratory manuals and practical recommendations. AB - CONTEXT: On-line clinical laboratory manuals are a valuable resource for medical professionals. To our knowledge, no recommendations currently exist for their content or design. OBJECTIVE: To analyze publicly accessible on-line clinical laboratory manuals and to propose guidelines for their content. DESIGN: We conducted an Internet search for clinical laboratory manuals written in English with individual test listings. Four individual test listings in each manual were evaluated for 16 data elements, including sample requirements, test methodology, units of measure, reference range, and critical values. Web sites were also evaluated for supplementary information and search functions. RESULTS: We identified 48 on-line laboratory manuals, including 24 academic or community hospital laboratories and 24 commercial or reference laboratories. All manuals had search engines and/or test indices. No single manual contained all 16 data elements evaluated. An average of 8.9 (56%) elements were present (range, 4-14). Basic sample requirements (specimen and volume needed) were the elements most commonly present (98% of manuals). The frequency of the remaining data elements varied from 10% to 90%. CONCLUSIONS: On-line clinical laboratory manuals originate from both hospital and commercial laboratories. While most manuals were user-friendly and contained adequate specimen-collection information, other important elements, such as reference ranges, were frequently absent. To ensure that clinical laboratory manuals are of maximal utility, we propose the following 13 data elements be included in individual test listings: test name, synonyms, test description, test methodology, sample requirements, volume requirements, collection guidelines, transport guidelines, units of measure, reference range, critical values, test availability, and date of latest revision. PMID- 15043485 TI - Steroid profiles using liquid chromatography-tandem mass spectrometry with atmospheric pressure photoionization source. PMID- 15043487 TI - [Inflammatory bowel disease: etio-pathogenic pieces to be sewn]. PMID- 15043486 TI - Balamuthia mandrillaris meningoencephalitis in an immunocompetent patient: an unusual clinical course and a favorable outcome. AB - Balamuthia mandrillaris meningoencephalitis is a rare but often fatal infection; only 2 survivors have been reported to date worldwide. We report the case of an apparently immunocompetent patient (72-year-old woman) who developed several episodes of seizures without prior history of respiratory or skin infections. Magnetic resonance imaging with contrast revealed 2 ring-enhancing lesions, one in the right precentral region and the other in the left posterotemporal region. Open biopsy revealed Balamuthia encephalitis. The patient was treated with combination antibiotics (pentamidine, 300 mg intravenously once a day; sulfadiazine, 1.5 g 4 times a day; fluconazole, 400 mg once a day; and clarithromycin, 500 mg 3 times a day) and was discharged home. There have been no significant neurological sequelae at this writing (6 months after biopsy). We present this case with unusual clinical course to raise awareness of this infectious disease, which may have a more favorable outcome if diagnosed and treated in its early states. PMID- 15043488 TI - [Subcutaneous panniculitis at a third level hospital retrospective study of 113 cases]. AB - INTRODUCTION: The panniculitides or hypodermitis are a wide and heterogeneous group of diseases characterized by the presence of subcutaneous inflammatory nodules located generally in low limbs. OBJECTIVES: Show the clinicopathologic characteristics of the cases diagnosed as panniculitis for a 5 years period; their epidemiology, etiopathogenic factors, evolution and treatment. MATERIAL AND METHODS: A retrospective study of 113 patients diagnosed as panniculitis by histopathologic report of cutaneous biopsy since 1997 to 2001 were performed. RESULTS: The sex proportion was of 4.65 women to every man. The mean age of these patients was 49 +/- 20 years old. 100% had legs affectation, 7.1% in arms, 5.9% in trunk and 0.9% in face. 9.7% presented fever and arthromyialgias, especially in young people (p < 0.05). The etiopathogenic factors implied were tuberculosis, streptococcic pharyngoamygdalitis, sarcoidosis and different drugs. The most frequent histological pattern was septal panniculitis without vasculitis. 24.8% did not realize any treatment; 32.5% took nonsteroideal anti-inflammatory drugs; 31.6% potassium iodide; 16% systemic corticosteroids and 7.1% tuberculostatic drugs. Evolution of the patients was good and independently from the treatment performed. CONCLUSIONS: Panniculitides are a group of diseases with similar clinic, but very heterogeneous in their etiology and histopathologic findings. They are more frequent in women and with more expressive clinic in young people. Up to in the 59.3% of the studied cases a causal etiologic agent was identified. These factors are compatible with the published ones in other previous panniculitis series. When a paniculitis is suspected, the most efficient tool is the clinical diagnosis. Histological confirmation by cutaneous biopsy it must be done when it is possible. PMID- 15043489 TI - [Cuttoff values of biochemical tests on pleural fluid: their usefulness in differential diagnosis of 1,040 patients with pleural effusion]. AB - OBJECTIVE: The aim of biochemical pleural fluid testing is to reach an etiological diagnosis of the pleural effusion. We assessed the utility of considering cuttoff points for the wide range of analytes used to investigate pleural fluid. PATIENTS AND METHODS: Among 1,040 patients with pleural effusion, we sought the etiologies of those fluids which showed any of the following characteristics: red blood cell count = 10 x 109/L, leukocytes = 10 x 109/L, percentage of neutrophils or lymphocytes >50%, protein = 50 g/L, glucose = 60 mg/dL, pH = 7.2, lactate dehydrogenase = 1,000 U/L, adenosine deaminase = 40 U/L, amylase = 100 U/L or cholesterol = 60 mg/dL. RESULTS: Some of the more prominent findings were: a) a sixth of transudates were blood-tinged or contained predominantly neutrophils; b) a groosly bloody fluid suggests malignant disease, trauma, or pulmonary embolization; c) nearly 90% of fluids containing = 10 x 10(9) leukocytes/L were parapneumonics; d) 73% of tuberculous pleural fluids had protein > or = 50 g/L, e) tuberculosis and parapneumonics explained more than 90% of fluids with high adenosine deaminase content; f) one third of amylase-rich pleural effusions were malignant; g) a low pleural glucose or pH levels indicate that patient probably has a parapneumonic, tuberculous or malignant etiology; y h) the diagnostic yield of pleural fluid cytology in malignant effusions was 57%, a percentage which raised to 94% in those with low glucose fluid level. CONCLUSIONS: Cuttof values of biochemical pleural fluid tests may greatly support particular causes of pleural effusions. PMID- 15043490 TI - [Risk of infections and glycemic control in type 2 diabetes mellitus]. AB - BACKGROUND AND OBJECTIVE: Although association between Diabetes Mellitus (DM) type 2 and prevalence of infections is a frequently considered issue in current literature there is few evidence of it. The objective of this study has been to analyse the infection risk in DM type 2 patients according to their glycemic control level (Hb Aic%). PATIENTS AND METHOD: In this cohort-study of 740 patients, we evaluated the incidence rates of infectious episodes in DM2-type diabetic patients during a period of 5 years, from May, 1997 to May, 2002. RESULTS: The average period of follow-up was 4.26 years. We found no significant correlation (p=0.33) between higher levels of HbAic% and presentation of infections. Data clearly show a direct correlation between risk of infection and number of associated chronic diseases (p=0.035), age (p=0.007) and diabetes duration (0.012). We also found an unexplained association between more aggressive treatment and higher glycemic levels (Hb Aic%) (p=0.0001) and also higher infection risk (0.038). CONCLUSIONS: Although there is no association between risk of infection and glycemic control (Hb Aic%) among analysed patients, this study shows a clear correlation between risk of infection and number of complications or number of associated chronic diseases. PMID- 15043491 TI - [Prevalence of cognitive decline in people older than 70 years admitted in an internal medicine service]. AB - BACKGROUND: Cognitive decline in older people is an important cause of burden for caregivers and assistance staff. It is a poor prognosis marker that favors the onset of different geriatric syndromes and increases the risk of institutionalization. As some causes of cognitive decline can be treated, its early diagnosis is fundamental for patients and caregivers. This, help us to design a care plan and a future follow up. The objective of this article is to measure the prevalence of Cognitive Decline before the onset of the acute disease that led patients older than 70 years, to an admission at the Internal Medicine Service. PATIENTS AND METHODS: A prospective randomized study was performed in the Internal Medicine Service of the Hospital de Mataro, which included 100 patients older than 70 years. Presence of Cognitive Decline was evaluated through the Blessed Scale during the previous month of the onset of the disease that motivated the admission. RESULTS: 43% of the patients had Cognitive Decline (42% male and 44% female). The prevalence of CD in patients older than 80 years was over 60%. CONCLUSIONS: Many of the admitted patients of an Internal Medicine Service older than 70 years had criteria for Cognitive Decline before the onset of the acute disease. Its diagnosis, study and therapeutic plan before the patient's discharge is a great opportunity to improve the quality of assistance. PMID- 15043492 TI - [Obstructive jaundice after cholecystectomy]. AB - In this article we present the case of a 72 year-old woman who three years after laparoscopic cholecystectomy develops obstructive jaundice. An MRI of the liver and biliary system revealed an hiliar mass that caused dilatation of the biliary tree. The patient underwent hepatic duct resection and reconstruction via hepaticojejunostomy. The histological examination of the surgical specimen identified an intramural biliary neuroma with no evidence of malignancy. PMID- 15043493 TI - [Paraneoplastic limbic encephalitis and lung cancer]. AB - Paraneoplastic limbic encephalitis (PLE) is a disorder characterized by severe cognitive dysfunction and seizures. It is usually associated with small cell lung carcinoma. Diagnosis is difficult because clinical markers are often lacking, and symptoms usually precede the diagnosis of cancer. Serological detection of antineuronal antibodies can be useful. We describe a patient with symptoms of limbic encephalitis, negative for paraneoplastic antibodies, in whom lung cancer was detected. PMID- 15043494 TI - [Haemophagocytic syndrome and cutaneous T-cell lymphoma]. AB - Haemophagocytic syndrome is a rare disorder of immune regulation. Clinical features are fever, haemophagocytosis, hepatosplenomegaly, pancytopenia, lymph nodes enlargement, hypertriglyceridaemia and coagulopathy. Systemic findings may also include pulmonary infiltrates, renal failure and an inappropriate antidiuretic state. It has been described associated to many diseases, like lymphomas, most of them T-cell lymphomas, infections and systemic diseases, among other entities. Haemopoietic cells are actively ingested by monocytes and macrophages in lymph nodes, bone marrow, liver and spleen. This is a diagnostic criteria for haemophagocytic syndrome. The treatment of haemophagocytic syndrome is difficult and requires intensive supportive therapy, treatment of coagulopathy disorders, high-dose corticosteroids and immunosuppressive treatment, although in most cases there is no response to treatment and the evolution is fatal. The T cell lymphomas that involve subcutaneous tissue and simulate panniculitis are classified recently as primary cutaneous T-cell lymphomas, and their presentation as inflammatory nodules of the legs are uncommon. PMID- 15043495 TI - [Parotiditis viral infection associated with Kikuchi-Fujimoto disease. Review of 1 case]. AB - Kikuchi-Fujimoto disease or necrotizing histiocytic lymphadenitis is a self limited process of lymphatic system that affects more frequently young women. Fever and adenopathy, often cervical, are typical. Diagnosis is fundamentally based on the affected lymph node biopsy, since laboratory data are often normal. It's a very rare entity in Spain, being more frequent in Oriental countries. It's a benign evolution process, with a spontaneous healing in some weeks or months. This is the reason because of this disease could be infradiagnosed. It's very important the differential diagnosis with malignant lymphomas, because Kikuchi Fujimoto disease doesn't need any treatment. PMID- 15043496 TI - [Olive oil: influence and benefits on some pathologies]. AB - The olive tree has been one of the agriculture bases in Mediterranean countries with a great economic and social significance. The oil derivative from it fruit can be classified in different kinds according with their quality, being the highest exponent the so-called pure olive oil that contribute in unquestionable benefits for the maintenance of health, illness prevention as well as a better evolution when the illness is present. There are some studies that prove these benefits in pathologies like cancer specially breast and stomach cancer (colon, endometrium and ovary cancer too). Gastrointestinal pathology like peptic ulcer, cholelithiasis and gastric mobility. Rheumatoid arthritis decreasing it development risk and improving it evolution. Diabetes mellitus increasing insulin sensibility and decreasing blood pressure and atherogenic lipoprotein. PMID- 15043497 TI - [Autoinflammatory disorders: a new concept in hereditary periodic fever syndromes]. AB - At last year the great scientific advances in genetics and molecular biology have led to a bigger knowledge about we nowadays call "Autoinflammatory syndromes", characterized by recurrent inflammatory episodes genetically determined and not mediated by autoimmunity. In this group, they are included the hereditary periodic fever syndromes: familial mediterranean fever (FMF), hyper Ig-D syndrome (HIDS), TNF-receptor-associated periodic syndrome (TRAPS), Muckle-Wells syndrome (MWS), familial cold autoinflammatory syndrome (FCAS), CINCA syndrome. The past 6 year have witnessed the identification of genes causing these diseases. Some of these genes encode proteins with a common domain (PYRIN domain). These protein are part of regulatory pathway of inflammation and apoptosis. The purpose of this article, is to carry out review of the genetic, clinical, molecular and rheumatologic aspect of these syndromes, in part unknown. Although they are not common, they are not absent in our diary clinical practise. Their study and research we will be able to obtain new knowledge that lead us to solve the complex inflammatory process. PMID- 15043498 TI - [Evolution of tetanus and levels of pseudocholinesterase]. PMID- 15043499 TI - [Chronic cough as presentation of subacute bacterial endocarditis]. PMID- 15043500 TI - [Colon adenocarcinoma found within an inguinal hernia sac]. PMID- 15043501 TI - [Bilateral hip fracture and myeloma]. PMID- 15043502 TI - [Neurologic symptoms as beginning of a thrombotic thrombocytopenic purpura]. PMID- 15043503 TI - [Streptococcus bovis septicemia and bifocal spondylodiscitis]. PMID- 15043504 TI - [Achilles bilateral tendonitis and levofloxacin]. PMID- 15043505 TI - [Celiac disease in a patient with Rome II criteria of irritable bowel syndrome]. PMID- 15043506 TI - [Unusual evolution of multinodular goiter: thyroid and parathyroid infiltration by amyloiditis]. PMID- 15043507 TI - Evidence for a complex risk profile in obese postmenopausal Turkish women with hypertriglyceridaemia and elevated apolipoprotein B. AB - The aim of the present study was to examine possible pathophysiological relationships among a wide array of proatherogenic risk factors in postmenopausal women. Fasting lipids, apoB (apolipoprotein B), BMI (body mass index) and waist circumference were measured in 178 women (59.4+/-7.2 years) from the Turkish Adult Risk Factor Study. Fasting levels of complement C3, insulin, SHBG (sex hormone-binding globulin), cortisol, oestradiol, testosterone and DHEA-S (dehydroepiandrosterone sulphate) were also determined. This is the first study to examine the relationships of all these variables with apoB. In the first of two different approaches, three groups of obese women were compared. Group 1 comprised women who were normolipidaemic with normal apoB; group 2, women who were hypertriglyceridaemic, but with normal apoB; and group 3, women who were hypertriglyceridaemic with elevated apoB. Complement C3, fasting insulin and glucose were significantly higher and HDL-C (high-density lipoprotein cholesterol) and SHBG levels were significantly lower in group 3 than in group 1. In the former group, the testosterone/SHBG ratio tended to be higher, indicating more free testosterone, than in group 1. The mean risk score in group 3 and the odds ratio for coronary disease by logistic regression analysis were significantly higher, 2.56 (confidence intervals, 1.12-5.85; P=0.026), compared with the two other groups combined. In examining the whole group, apoB levels correlated significantly with a wider array of pro-atherogenic risk factors than did LDL-C (low-density lipoprotein-cholesterol), particularly being linked to complement C3 and glucose, as well as the risk score. Complement C3 demonstrated the widest associations and was significantly linked with BMI, waist circumference, insulin, glucose, fibrinogen, triacylglycerols (triglycerides) and apoB and was inversely correlated with HDL-C and SHBG. SHBG was also correlated inversely with a wide spectrum of risk variables. In summary, in Turkish women, apoB was linked with a complex array of proatherogenic risk factors, and hypertriglyceridaemia with elevated apoB was associated with a higher risk of coronary disease. PMID- 15043508 TI - Plasma expansion by polyethylene-glycol-modified albumin. AB - Systemic inflammatory response conditions are associated with capillary leak and haemodynamic compromise. Fluid resuscitation to reverse the ensuing hypovolaemia is, however, complicated by the decreased endothelium reflection coefficient to albumin and other colloids. We developed PEG-Alb (albumin covalently linked to polyethylene glycol) as a potential resuscitative agent. PEG was covalently linked to human albumin at multiple sites on the protein. The modified protein was heterogeneous when examined by SDS/PAGE, size-exclusion chromatography and SELDI-TOF MS (surface-enhanced laser-desorption ionization-time of flight MS). Based on size-exclusion chromatography and osmotic pressure data, the effective volume of PEG-Alb is increased 13- to 16-fold compared with unmodified albumin. In an LPS (lipopolysaccharide) model of shock, rats treated with PEG-Alb showed better blood pressure, lower Hct (haematocrit) consistent with haemodilution and less lung injury than rats treated with unmodified albumin or saline. In a CLP (caecal ligation and puncture) model of sepsis, PEG-Alb was more effective than albumin or saline in maintaining blood pressure and in decreasing Hct. When fluorescein-labelled PEG-Alb and Texas Red-labelled albumin were administered to rats with LPS- or CLP-induced shock, PEG-Alb was retained within blood vessels, whereas albumin extravasates into the interstitial space. Based on these data, PEG-Alb appears to be retained within blood vessels in models of capillary leak. PEG-Alb may ultimately be effective in the clinical treatment of shock associated with capillary leak. PMID- 15043509 TI - Probucol aggravates long QT syndrome associated with a novel missense mutation M124T in the N-terminus of HERG. AB - Patients with LQTS (long QT syndrome) with a mutation in a cardiac ion channel gene, leading to mild-to-moderate channel dysfunction, may manifest marked QT prolongation or torsade de pointes only upon an additional stressor. A 59-year old woman had marked QT prolongation and repeated torsade de pointes 3 months after initiation of probucol, a cholesterol-lowering drug. We identified a single base substitution in the HERG gene by genetic analysis. This novel missense mutation is predicted to cause an amino acid substitution of Met(124)-->Thr (M124T) in the N-terminus. Three other relatives with this mutation also had QT prolongation and one of them had a prolonged QT interval and torsade de pointes accompanied by syncope after taking probucol. We expressed wild-type HERG and HERG with M124T in Xenopus oocytes and characterized the electrophysiological properties of these HERG channels and the action of probucol on the channels. Injection of the M124T mutant cRNA into Xenopus oocytes resulted in expression of functional channels with markedly smaller amplitude. In both HERG channels, probucol decreased the amplitude of the HERG tail current, decelerated the rate of channel activation, accelerated the rate of channel deactivation and shifted the reversal potential to a more positive value. The electrophysiological study indicated that QT lengthening and cardiac arrhythmia in the two present patients were due to inhibition of I(Kr) (rapidly activating delayed rectifier K(+) current) by probucol, in addition to the significant suppression of HERG current in HERG channels with the M124T mutation. PMID- 15043510 TI - Purification and biochemical characterization of a thermostable, alkaliphilic, extracellular alpha-amylase from Bacillus subtilis DM-03, a strain isolated from the traditional fermented food of India. AB - Bacillus subtilis strain DM-03, which is isolated from starter culture used for the production of alcohol by local Assam tribes, grows optimally at 52-55 degrees C and secretes a significant amount of alpha-amylase at pH 8.0 into the culture media. This alpha-amylase, purified by ion-exchange, gel-filtration and reverse phase HPLC, constitutes 2.9% of the total extracellular protein. This purified enzyme, named Bsamy-I, has a subunit with molecular mass of 42.8 kDa as determined by SDS/PAGE, and optimum temperature and pH values at 52-55 degrees C and 9.0 respectively, which makes it ideal for use in the detergent industries. Maximum alpha-amylase production is obtained by using soluble starch and NH(4)Cl as carbon and nitrogen sources respectively. Thermostability of the enzyme is evident from heating the enzyme at 95 degrees C for 10 min, which results in a loss of 60% of the original enzyme activity. 4-Bromophenacyl bromide and PMSF at 4 and 1.5 mM concentration respectively completely abolish the enzymic activity, documenting the essential role of histidine and carboxylic residues in the catalytic process. PMID- 15043511 TI - Review article: current management of alcoholic liver disease. AB - Alcoholic liver disease, including acute alcoholic hepatitis and alcoholic cirrhosis, is a major cause of morbidity and mortality in the Western world. Abstinence remains the cornerstone of management of all forms of alcoholic liver disease. Recent research, which has elucidated the mechanisms of alcohol-induced liver injury, offers the prospect of advances in the management of alcoholic liver disease. We review the most recent data on the efficacy of treatment of acute alcoholic injury, including nutritional support, corticosteroids, anti inflammatory agents and antioxidants, and agents that are directed against the progression to fibrosis, such as colchicines, propylthiouracil and antioxidants. Although these therapies offer a tantalizing glimpse into a future that may include therapies that directly alter the process of injury and repair in the liver, none has been shown consistently to improve the course of alcoholic liver damage. Consequently, liver transplantation remains an ultimate option for selected patients with liver failure due to chronic alcoholic liver damage. PMID- 15043512 TI - Review article: hepatitis vaccination in patients with chronic liver disease. AB - Evidence regarding the outcomes of viral super-infection in patients with chronic liver disease and practical strategies for hepatitis A and B vaccination of these individuals are reviewed. Patients with acute hepatitis A and chronic hepatitis B have a more severe clinical course and a higher death rate compared with otherwise healthy individuals with hepatitis A, and these differences are most pronounced in older patients and those with histological evidence of chronic hepatitis or cirrhosis, rather than in asymptomatic hepatitis B carriers. Patients with acute hepatitis A super-infection and chronic hepatitis C have an increased risk of fulminant hepatitis and death. In addition, patients with other chronic liver diseases also appear to be at increased risk for more severe disease with superimposed hepatitis A. Patients with chronic hepatitis B and hepatitis C virus co-infection have more severe laboratory abnormalities, more severe histological disease, a greater frequency of cirrhosis and complications of cirrhosis, and a higher incidence of hepatocellular carcinoma. Vaccines for both hepatitis A and B are safe and effective if used early in the course of chronic liver disease. Hepatitis A and B vaccination should be part of the routine management of patients with chronic liver disease, preferably as early as possible in the natural course of their disease. PMID- 15043513 TI - Systematic review: the epidemiology of ischaemic colitis. AB - BACKGROUND: Ischaemic colitis has been associated with co-morbid conditions, medications, vascular surgery and advanced age in case reports and case series. Few data exist on the baseline incidence in the general population or on the increased risk imposed by these risk factors. AIM: To systematically review the literature regarding the incidence, prevalence and risk factors for ischaemic colitis. METHODS: Searches of bibliographic databases were performed independently by two investigators. Studies were included if they used population based samples, disease-specific population samples or case-control population based samples of adults with ischaemic colitis, and reported the incidence, prevalence or risk factors for ischaemic colitis. Eligible articles were reviewed and data were abstracted in a duplicate, independent manner. RESULTS: Four studies were identified that reported the general population incidence and four that reported the disease-specific population incidence. The incidence of ischaemic colitis in general populations ranged from 4.5 to 44 cases per 100 000 person-years. The risk was increased two- to four-fold by either prevalent irritable bowel syndrome or chronic obstructive pulmonary disease. The risk was also increased in females and in subjects of 65 years and older. CONCLUSIONS: Ischaemic colitis is uncommon in the general population. The effect sizes of the most commonly reported risk factors have not been adequately quantified in population-based studies. PMID- 15043514 TI - Randomized, double-blind, placebo-controlled trial of oral aloe vera gel for active ulcerative colitis. AB - BACKGROUND: The herbal preparation, aloe vera, has been claimed to have anti inflammatory effects and, despite a lack of evidence of its therapeutic efficacy, is widely used by patients with inflammatory bowel disease. AIM: To perform a double-blind, randomized, placebo-controlled trial of the efficacy and safety of aloe vera gel for the treatment of mildly to moderately active ulcerative colitis. METHODS: Forty-four evaluable hospital out-patients were randomly given oral aloe vera gel or placebo, 100 mL twice daily for 4 weeks, in a 2 : 1 ratio. The primary outcome measures were clinical remission (Simple Clinical Colitis Activity Index /= 3 points; response was defined as remission or improvement), Baron score, histology score, haemoglobin, platelet count, erythrocyte sedimentation rate, C-reactive protein and albumin. RESULTS: Clinical remission, improvement and response occurred in nine (30%), 11 (37%) and 14 (47%), respectively, of 30 patients given aloe vera, compared with one (7%) [P = 0.09; odds ratio, 5.6 (0.6-49)], one (7%) [P = 0.06; odds ratio, 7.5 (0.9-66)] and two (14%) [P < 0.05; odds ratio, 5.3 (1.0-27)], respectively, of 14 patients taking placebo. The Simple Clinical Colitis Activity Index and histological scores decreased significantly during treatment with aloe vera (P = 0.01 and P = 0.03, respectively), but not with placebo. Sigmoidoscopic scores and laboratory variables showed no significant differences between aloe vera and placebo. Adverse events were minor and similar in both groups of patients. CONCLUSION: Oral aloe vera taken for 4 weeks produced a clinical response more often than placebo; it also reduced the histological disease activity and appeared to be safe. Further evaluation of the therapeutic potential of aloe vera gel in inflammatory bowel disease is needed. PMID- 15043515 TI - The risk of post-operative complications associated with infliximab therapy for Crohn's disease: a controlled cohort study. AB - BACKGROUND: By temporarily suppressing the immune response, the anti-tumour necrosis factor agent, infliximab, may increase the risk of peri-operative complications. AIM: To test this hypothesis for intestinal resection in a cohort of 313 Crohn's disease patients treated with infliximab. Forty received one or more infusions prior to intestinal resection (31/40 within 12 weeks). METHODS: The post-operative events of these patients were compared with those of a control group (infliximab naive) of 39 patients adjusted for age, gender and surgical procedure. Early (10 days) and late (3 months) major or minor complications were identified. RESULTS: The incidence of early minor (15.0% vs. 12.8%) and major (12.5% vs. 7.7%) and late minor (2.5% vs. 5.1%) and major (17.5% vs. 12.8%) complications and the mean hospital stay after surgery (10.3 +/- 4.0 days vs. 9.9 +/- 5.5 days) were similar in both groups. A trend towards an increased early infection rate was found in infliximab pre-treated patients (6 vs. 1; P = 0.10), but more patients in this group received corticosteroids and/or immunosuppressives (29 vs. 16 patients; P < 0.05). CONCLUSION: The use of infliximab before intestinal resection does not prolong the hospital stay and does not increase the rate of post-operative complications. PMID- 15043516 TI - Rofecoxib and early relapse of inflammatory bowel disease: an open-label trial. AB - BACKGROUND: The safety and efficacy of selective cyclo-oxygenase-2 inhibitors in inflammatory bowel disease are under investigation. AIM: To assess, in a prospective, open-label trial, the efficacy and safety of rofecoxib (12.5 mg/day) in inflammatory bowel disease patients and controls. METHODS: The inflammatory bowel disease group included 45 inactive patients (25 Crohn's disease; 20 ulcerative colitis) with associated arthralgia. The control group included 30 dyspeptic patients. The efficacy and safety of rofecoxib were assessed in inflammatory bowel disease patients and controls before and after treatment (range, 3 days to 3 months). RESULTS: In inflammatory bowel disease, nine of the 45 patients (20%) required rofecoxib withdrawal due to gastrointestinal symptoms inducing clinical relapse, which subsided on drug discontinuation. The percentage of patients requiring rofecoxib discontinuation was comparable in patients with Crohn's disease and ulcerative colitis (20% vs. 20%), but was higher in inflammatory bowel disease patients than in controls (20% vs. 3%; P < 0.001). In inflammatory bowel disease, arthralgia relief was reported by 32 patients (71%): complete relief by eight patients (18%) and partial relief by 24 (53%). Thirteen patients (29%) reported no benefit. A comparable percentage of inflammatory bowel disease patients and controls reported arthralgia relief (71% vs. 70%). CONCLUSIONS: Rofecoxib appears to control arthralgia in almost two-thirds of inflammatory bowel disease patients. Side-effects requiring drug discontinuation are observed, however, in almost one-quarter of patients. PMID- 15043517 TI - Diverticular disease of the colon: ethnic differences in frequency. AB - BACKGROUND: Colonic diverticular disease is more common in Western populations than in developing countries. AIM: To determine whether the frequency of colonic diverticular disease is different in British patients of Indian-subcontinent Asian origin compared with other ethnic groups. METHODS: All colonoscopies performed over a 3-year period in a London hospital were studied. Patients of Indian-subcontinent Asian origin were identified by name. RESULTS: Five of 134 Indian-subcontinent Asian males (4%) had colonic diverticular disease, compared with 278 of 1268 patients of other ethnic groups (22%; P < 0.001). Five of 91 Indian-subcontinent Asian females (6%) had colonic diverticular disease, compared with 333 of 1486 patients of other ethnic groups (23%; P < 0.001). Although patients of Indian-subcontinent Asian origin (54.8 +/- 15.8 years) were younger than those of other ethnic groups (60.3 +/- 17.8 years; P < 0.0001), the ethnic difference in the frequency of diverticular disease persisted even when age was taken into account. CONCLUSION: There is a lower frequency of colonic diverticular disease in Indian-subcontinent Asians presenting for colonoscopy, compared with other ethnic groups. This cannot be explained by sex or age differences. Our findings require confirmation, but may provide opportunities for research into the aetiology of colonic diverticular disease. PMID- 15043518 TI - Different presentation of hepatitis B-related hepatocellular carcinoma in a cohort of 1863 young and old patients - implications for screening. AB - AIM: To compare the clinico-pathological features of hepatitis B virus-related hepatocellular carcinoma in young and old patients. METHODS: The clinico pathological characteristics of hepatitis B virus-related hepatocellular carcinoma were compared in 1863 consecutive patients (121 patients, 40 years) seen at a single institution over the last 13 years. RESULTS: Young patients presented more often with pain (P < 0.0001), hepatomegaly (P = 0.01) and ruptured hepatocellular carcinoma (P = 0.02), whereas old patients presented with ankle oedema (P = 0.001), ascites (P = 0.002) and by routine screening (P = 0.035). Liver function, Child-Pugh grading and indocyanine green test were better preserved in young patients. They also had a higher alpha foetoprotein concentration (P = 0.001), larger tumour size (P = 0.001) and more frequent metastasis (P = 0.008), but a similar surgical resection rate (33.6% vs. 28%), to old patients. There was no difference between the two groups in the overall post-resection survival rate, but there was a shorter survival in young patients with unresectable disease (3.6 months vs. 4.6 months, P = 0.004). Young patients with hepatocellular carcinoma often show a later presentation, but a higher resectability rate and similar survival rates, than old patients. The screening programme should include young hepatitis B virus carriers, even in the absence of cirrhosis. PMID- 15043519 TI - Long-term oral branched chain amino acids in patients undergoing chemoembolization for hepatocellular carcinoma: a randomized trial. AB - BACKGROUND: Patients undergoing transarterial chemoembolization for hepatocellular carcinoma have advanced tumour or severe cirrhosis and frequently have associated protein-calorie malnutrition. The role of nutritional supplements for such patients is unclear. AIM: To investigate, in a randomized controlled trial, any benefit of the long-term administration of branched chain amino acids in patients undergoing chemoembolization for hepatocellular carcinoma. METHODS: Forty-one patients received oral branched chain amino acids for up to four courses of chemoembolization and 43 patients did not receive any nutritional supplement. Morbidity, liver function, nutritional status, quality of life and long-term survival were compared between the two groups. RESULTS: The administration of branched chain amino acids resulted in a lower morbidity rate compared with the control group (17.1% vs. 37.2%, P = 0.039). In particular, the group given branched chain amino acids showed a significantly lower rate of ascites (7.3% vs. 23.2%, P = 0.043) and peripheral oedema (9.8% vs. 27.9%, P = 0.034). Significantly higher serum albumin, lower bilirubin and a better quality of life were observed after chemoembolization in the group given branched chain amino acids. However, there was no significant difference in survival between the two groups. CONCLUSIONS: Nutritional supplementation with oral branched chain amino acids is beneficial in increasing the serum albumin level, reducing the morbidity and improving the quality of life in patients undergoing chemoembolization for inoperable hepatocellular carcinoma. PMID- 15043520 TI - High efficacy of 1-week doxycycline- and amoxicillin-based quadruple regimen in a culture-guided, third-line treatment approach for Helicobacter pylori infection. AB - BACKGROUND: Helicobacter pylori infection may persist after both first- and second-line current treatments. AIM: To assess the efficacy of a third-line, culture-guided treatment approach for the eradication of H. pylori. METHODS: Patterns of resistance were analysed in H. pylori isolates from 94 consecutive patients in whom H. pylori infection had persisted after two eradication attempts. Using the epsilometer test, susceptibility analysis was performed for amoxicillin, clarithromycin, metronidazole, tetracycline and levofloxacin. Patients were then treated with a culture-guided, third-line regimen: 89 patients with a 1-week quadruple regimen including omeprazole, bismuth, doxycycline and amoxicillin, and five patients with a 1-week triple regimen containing omeprazole, amoxicillin and levofloxacin or clarithromycin. RESULTS: Ninety-four subjects (100%) were resistant to metronidazole, 89 (95%) to clarithromycin, 29 (31%) to levofloxacin and five (5%) to tetracycline. No resistance to amoxicillin was found in any patient. Overall, H. pylori eradication was obtained in 90% of subjects. The quadruple regimen was effective in 81 patients (92% by per protocol and 91% by intention-to-treat analysis). Four patients (80%, both per protocol and intention-to-treat analysis) were H. pylori-negative after the triple regimen. CONCLUSIONS: A culture-guided, third-line therapeutic approach is effective for the eradication of H. pylori. Furthermore, the 1-week doxycycline- and amoxicillin-based quadruple regimen is a good third-line 'rescue' treatment option. PMID- 15043521 TI - Gallbladder volume as a biomarker for the motilin effect in healthy volunteers and patients with functional dyspepsia. AB - AIM: To investigate a motilin effect on gallbladder volume in healthy volunteers and patients with functional dyspepsia. METHODS: Forty-three healthy volunteers and 10 patients with functional dyspepsia received motilin (4 pmol.min/kg) or placebo in four separate double-blind, randomized, placebo-controlled, cross-over studies. The gallbladder volume was measured by ultrasonography. Analysis of variance of the combined data of these studies was performed to investigate a motilin effect on gallbladder volume and potential differences between patients and healthy volunteers. RESULTS: The baseline gallbladder volume was similar for placebo and motilin treatment, as well as for patients and healthy volunteers. Motilin, compared with placebo, significantly decreased the gallbladder volume in healthy volunteers (P = 0.003) and patients (P < 0.0001). A linear concentration response relationship was observed. The decrease in gallbladder volume by motilin was greater in patients (P = 0.03). The motilin effect was consistent between studies. CONCLUSION: The interdigestive gallbladder volume is a non-invasive end point for motilin activity, displaying a consistent response across studies, a clear response to motilin and a clear concentration-response relationship. However, it is less suitable as a biomarker for future pharmacological studies on motilin agonists or antagonists as the effect is probably indirect, and a relatively large study population of 27 subjects is required to demonstrate a 15% decrease in gallbladder volume. Further investigation is required to confirm altered gallbladder motility as a feature of functional dyspepsia. PMID- 15043522 TI - Inter-digestive and post-prandial antro-pyloro-duodenal motor activity in humans: effect of 5-hydroxytryptamine 1 receptor agonism. AB - BACKGROUND: Little is known about the effect of 5-hydroxytryptamine 1 (5-HT(1)) receptor agonism on the co-ordinated motor activity of the gastric antrum, pylorus and duodenum under fasting and fed conditions. AIM: To evaluate the effect of sumatriptan, a 5-HT(1) agonist, on fasting and fed antro-pyloro duodenal motility. METHODS: In study 1, antro-pyloro-duodenal motility was recorded for two phase IIIs of the migrating motor complex and then, following either a subcutaneous injection of sumatriptan 6 mg or saline control, for at least one additional phase III in 11 healthy volunteers (21-36 years). In study 2, the post-prandial motility was recorded for 3 h after either a subcutaneous injection of sumatriptan 6 mg or saline control in 10 healthy volunteers (18-36 years). RESULTS: Sumatriptan prolonged the migrating motor complex cycle (P = 0.009) by increasing the duration of phase II (P = 0.02) but not phases I and III. Post-prandially, sumatriptan reduced the activity index (P = 0.017) by reducing the frequency of co-ordinated motor activity involving the antrum and/or the duodenum (P < 0.05). CONCLUSION: 5-HT(1) receptor agonism increases the periodicity of the migrating motor complex and reduces the occurrence of post prandial co-ordinated motor activity involving the gastric antrum, pylorus and duodenum. PMID- 15043523 TI - COX-2-selective inhibitors and the risk of upper gastrointestinal bleeding in high-risk patients with previous gastrointestinal diseases: a population-based case-control study. AB - BACKGROUND: Clinical trials have suggested that cyclo-oxygenase-2-selective inhibitors are associated with a lower risk of upper gastrointestinal bleeding than are non-selective, non-aspirin, non-steroidal anti-inflammatory drugs. This has not yet been confirmed in studies of patients with an increased susceptibility to upper gastrointestinal bleeding. AIM: To examine the risk of upper gastrointestinal bleeding in high-risk patients who filled prescriptions for cyclo-oxygenase-2 inhibitors or other non-steroidal anti-inflammatory drugs. METHODS: A population-based case-control study was performed in the Danish county of North Jutland from 1 January 2000 to 31 December 2002. From the County Hospital Discharge Registry and the Civil Registration System, we identified incident cases with upper gastrointestinal bleeding (n = 780) and randomly selected controls (n = 2906), respectively. All cases and controls had previous gastrointestinal diseases. Data on drug exposure were obtained from the countywide Prescription Database. RESULTS: Thirty-five cases (4.5%) filled prescriptions for cyclo-oxygenase-2 inhibitors within 30 days of the date of upper gastrointestinal bleeding, compared with 79 controls (2.7%). Adjusted odds ratios for upper gastrointestinal bleeding according to prescription for celecoxib, rofecoxib and non-steroidal anti-inflammatory drugs were 1.3 [95% confidence interval (CI), 0.7-2.8], 2.1 (95% CI, 1.2-3.5) and 3.3 (95% CI, 2.4 4.4), respectively. CONCLUSIONS: In patients with increased susceptibility to gastrointestinal adverse events, a lower risk of upper gastrointestinal bleeding was observed in users of cyclo-oxygenase-2 inhibitors compared with users of other non-aspirin, non-steroidal anti-inflammatory drugs. PMID- 15043524 TI - Non-invasive assessment of liver function: trying not to miss an opportunity. PMID- 15043526 TI - Are there advantages in implanting a yellow IOL to reduce the risk of AMD? PMID- 15043527 TI - Sensitivity and specificity of digital retinal images in grading diabetic retinopathy. AB - PURPOSE: To assess three novel digital fundus cameras for diabetic retinopathy (DR) screening. METHODS: Digital colour and red-free retinal imaging (Topcon TRC 50 IA, Canon CR6-45NM, and MediTell) was used to capture 427 images of 70 diabetes patients and control subjects. The images were graded for DR by three readers in a randomized and masked manner using a modified Early Treatment Diabetic Retinopathy Study classification. The reference standard was based on mydriatic ophthalmoscopy and colour and red-free images. RESULTS: Digital 50 degrees red-free imaging had sensitivity of 97.7%, two-field 50 degrees colour imaging 94.0%, and two-field 45 degrees colour imaging sensitivity of 88.9%. The specificity of these imaging modalities was 98.9-100%, and ungradeable images represented 1.2-1.6%. The hand-held digital colour videocamera (MediTell) showed a sensitivity of 6.9% and ungradeable images represented 92.3%. CONCLUSION: Digital 50 degrees red-free and two-field 50 degrees or 45 degrees colour imaging were suitable for DR screening, whereas the hand-held digital videocamera did not fulfil the needs of DR screening. PMID- 15043528 TI - Evaluation of visual impairment in Usher syndrome 1b and Usher syndrome 2a. AB - PURPOSE: To evaluate visual impairment in Usher syndrome 1b (USH1b) and Usher syndrome 2a (USH2a). METHODS: We carried out a retrospective study of 19 USH1b patients and 40 USH2a patients. Cross-sectional regression analyses of the functional acuity score (FAS), functional field score (FFS) and functional vision score (FVS) related to age were performed. Statistical tests relating to regression lines and Student's t-test were used to compare between (sub)groups of patients. Parts of the available individual longitudinal data were used to obtain individual estimates of progressive deterioration and compare these to those obtained with cross-sectional analysis. Results were compared between subgroups of USH2a patients pertaining to combinations of different types of mutations. RESULTS: Cross-sectional analyses revealed significant deterioration of the FAS (0.7% per year), FFS (1.0% per year) and FVS (1.5% per year) with advancing age in both patient groups, without a significant difference between the USH1b and USH2a patients. Individual estimates of the deterioration rates were substantially and significantly higher than the cross-sectional estimates in some USH2a cases, including values of about 5% per year (or even higher) for the FAS (age 35-50 years), 3-4% per year for the FFS and 4-5% per year for the FVS (age > 20 years). There was no difference in functional vision score behaviour detected between subgroups of patients pertaining to different biallelic combinations of specific types of mutations. CONCLUSIONS: The FAS, FFS and FVS deteriorated significantly by 0.7-1.5% per year according to cross-sectional linear regression analysis in both USH1b and USH2a patients. Higher deterioration rates (3-5% per year) in any of these scores were attained, according to longitudinal data collected from individual USH2a patients. Score behaviour was similar across the patient groups and across different biallelic combinations of various types of mutations. However, more elaborate studies, preferably covering longitudinal data, are needed to obtain conclusive evidence. PMID- 15043529 TI - Activated protein C resistance in anterior ischaemic optic neuropathy. AB - PURPOSE: Protein C is a major component of the natural anticoagulant pathway. Resistance of coagulation factor V (FV) to activated protein C (APC), mostly due to FV Leiden mutation, is the most common cause of inherited thrombophilia. The aim of this retrospective study was to determine the prevalence of APC resistance and Leiden mutation in patients with non-arteritic anterior ischaemic optic neuropathy (NAION). METHODS: A total of 25 patients with NAION were examined between 1997 and 2002. The patients were screened for APC resistance and FV Leiden mutation as well as for acquired risk factors of vascular disease such as diabetes mellitus, hypercholesterolaemia, hypertonia and ischaemic heart disease. A control group of subjects without ocular vascular disease and with homogenous distribution of the same risk factors was used for comparison. RESULTS: Six of the 25 patients (24%) with NAION had APC resistance due to the heterozygous Leiden mutation of FV. The frequency of the same genetic mutation in the control group was only 5.9%. Odds ratio calculations showed that patients with the Leiden mutation were at a significantly higher risk of NAION than control patients (p < or = 0.021). CONCLUSION: The high frequency of Leiden mutation in NAION suggests a pathogenic role of the mutation in the disease. PMID- 15043531 TI - Phototherapeutic keratectomy in Salzmann's nodular degeneration. AB - PURPOSE: To describe the outcome in vision and refraction of phototherapeutic keratectomy (PTK) in Salzmann's nodular degeneration (SND). METHODS: Five eyes of four patients underwent PTK with the objective of restoring an acceptable refractive status and improving visual acuity. This surgical technique aims to make the nodules level with the corneal surface using an excimer laser. RESULTS: Best corrected visual acuity (BCVA) improved in all five eyes. Astigmatism was reduced in four eyes and increased somewhat in one. During the observation period one eye suffered a late recurrence that required a penetrating graft. CONCLUSION: Salzmann's nodular degeneration is a rare disease that is sometimes difficult to diagnose as it has some resemblance to other diseases. The aetiology of the disease is unknown. Phototherapeutic keratectomy is a safe and effective mode of treatment. Recurrences occur with time. PMID- 15043530 TI - Evaluation of surgical performance with intracameral mydriatics in phacoemulsification surgery. AB - PURPOSE: To evaluate surgical performance using intracameral mydriatics (ICM) in phacoemulsification surgery in a series of consecutive cases. METHODS: In a series of 198 consecutive procedures 50-200 microl of cyclopentolate 0.1%, phenylephrine 1.5% and lidocaine 1% was given intracamerally for mydriasis and anaesthesia. The previous 198 cases, dilated with topical mydriatics, were studied for comparison. Several pre- intra- and postoperative parameters were registered, and the subjective surgical performance was graded after each procedure. In 41 consecutive cases, the change in pulse and oxygen saturation induced by the ICM injection was registered. RESULTS: No increase in operation time or complication rates was seen with ICM, compared to when standard topical mydriatics were used. The subjective surgical performance was ranked as equally good for both groups. CONCLUSION: From this clinical evaluation, our impression is that ICM performs well in routine phacoemulsification surgery. PMID- 15043532 TI - Gonioscopic view of the trabeculo-Descemet's membrane after deep sclerectomy. AB - PURPOSE: To study the appearance of the trabeculo-Descemet's membrane (TDM) after deep sclerectomy with collagen implant (DSCI). METHODS: All patients who underwent a successful DSCI for open-angle glaucoma were included in a non randomized clinical study. Gonioscopy was performed between 6 and 12 months postoperatively in all patients. RESULTS: Gonioscopy was performed in 42 eyes of 38 patients after a mean follow-up time of 8.3 +/- 2.5 months after DSCI. The diagnosis was chronic open-angle glaucoma in 16 eyes and exfoliative glaucoma in 24 eyes. One patient had pigmentary glaucoma and one had normal tension glaucoma. None of the patients were on glaucoma medication postoperatively. Synechias to the TDM were seen in five eyes. Pigment accumulation to the TDM was seen in nine eyes, four of which had capsular glaucoma, three had primary open-angle glaucoma, one had pigmentary glaucoma and one had normal tension glaucoma. Puncture holes at the TDM were seen in all eight eyes in which Nd:YAG laser goniopuncture had been performed. CONCLUSION: Postoperative changes, which may go unnoticed without routine gonioscopy, occur at the TDM after deep sclerectomy with collagen implant. Pigment accumulates relatively shortly after surgery. Whether or not these changes are progressive remains unknown. Careful follow-up after deep sclerectomy is necessary and should include routine gonioscopy. PMID- 15043533 TI - Assessment of chamber angle pigmentation during longterm latanoprost treatment for open-angle glaucoma. AB - PURPOSE: This study aimed to determine whether the longterm use of latanoprost is associated with an increase in trabecular pigmentation, especially in subjects in whom iris pigmentation has increased. METHODS: We enrolled 50 subjects for whom treatment was to start for ocular hypertension, primary open-angle glaucoma or normal tension glaucoma. All subjects received latanoprost 0.005% daily. Trabecular pigmentation was documented using gonioscopic photography of the inferior quadrant at baseline, every 3 months for the first year and every 6 months for the second and third years. Three glaucoma specialists evaluated the series of gonioscopic photographs for each eye of each subject in a masked fashion. The intraocular pressure (IOP) was also recorded at each visit. RESULTS: A total of 41 subjects (79 eyes) completed 3 years of follow-up, and none showed any increase in the grade of trabecular pigmentation, including 10 subjects (20 eyes) in whom the iridial pigment increased. CONCLUSION: Although latanoprost increased iridial pigmentation in some subjects, we found no evidence of an increase in trabecular pigmentation over the 3 years of follow-up. PMID- 15043534 TI - The safety and efficacy of unoprostone 0.15% versus brimonidine 0.2%. AB - PURPOSE: To compare the efficacy and safety of unoprostone versus brimonidine both given twice daily in ocular hypertensive or primary open-angle glaucoma subjects. METHODS: After a 1-month washout period a baseline diurnal curve was measured every 2 hours from 08:00 hours (trough) to 20:00 hours in subjects with a trough intraocular pressure (IOP) and the pressure 24 mmHg. Qualified subjects were randomized to either brimonidine or unoprostone. After 6 weeks of treatment the period 1 diurnal curve was performed. Subjects were then switched to the opposite treatment for 6 weeks and the period 2 diurnal curve was performed. RESULTS: A total of 33 subjects were included in this study. In the brimonidine treated group the trough IOP 20.1 +/- 2.8 mmHg was reduced from baseline up to 8 hours after dosing. In the unoprostone-treated group the trough IOP was 19.5 +/- 3.0 mmHg, which was statistically equal to that of brimonidine (p = 0.21), was reduced from baseline for 12 hours after dosing. Brimonidine decreased the IOP statistically more than unoprostone at 10:00 and 12:00 hours (p < 0.0001 and p = 0.02, respectively), while unoprostone reduced the IOP more than brimonidine at 18:00 and 20:00 hours (p = 0.002 and p = 0.05, respectively). Safety levels were similar between groups, but unoprostone caused more ocular stinging than brimonidine (p = 0.008). CONCLUSION: This study suggests that twice daily brimonidine demonstrates a statistically greater peak reduction in IOP than unoprostone. However, unoprostone, but not brimonidine, decreased IOP over the complete 12-hour daytime dosing cycle. PMID- 15043535 TI - Ocular changes, risk markers for eye disorders and effects of cataract surgery in elderly people: a study of an urban Swedish population followed from 70 to 97 years of age. AB - AIMS: To investigate the prevalence of and potential risk factors for ocular disorders and the effects of timing of cataract surgery from age 70-97 years. POPULATION: A representative population sample taken from within the Gerontological and Geriatric Population Studies (H 70) in Gothenburg, Sweden (n = 958). All subjects underwent eye examinations at age 70 years in 1971 and subsequently at ages 82, 88, 95 and 97 years. All inhabitants of Gothenburg aged 95 and 97 years were invited to participate in the study. RESULTS: Decreased vision (visual acuity < or = 0.5) was found in 20% and 80% of subjects at ages 82 and 97 years, respectively. Blood folate and physical activity at age 70 years correlated positively and body mass index (BMI) negatively to visual acuity (VA) > or = 0.8 at ages 82 and 88 years. Smoking at age 70 years correlated to early age-related maculopathy (ARM). Cataract surgery had been performed in 40% of subjects at age 97 years. Surgery 2 years earlier led to a 15% increase in time spent with improved vision. CONCLUSIONS: The deterioration of vision in elderly people is a major health problem, for which 'low' folate status, smoking, 'high' BMI and low physical activity are potential risk factors. Early cataract surgery is also beneficial in very old patients. PMID- 15043536 TI - Ocular anisometropia and laterality. AB - PURPOSE: To study the difference in refraction between right and left eyes (anisometropia) in different age groups, look for evidence of eye laterality (more refractive error in one eye) and compare the size of anisometropia in the myopic and emmetropic ranges. METHODS: The study was based on children in Hong Kong (examined at the age of 6 years and again at the age of 8.5 years) and their parents (aged 26-60 years). RESULTS: In all age groups the difference between right and left eyes in sphere and cylinder was modest, in most cases < or = 0.25 D. In both children and their parents a tendency towards spherical right eye laterality was found in the myopic individuals (p < 0.05). This was not the case with the cylinder. In the 8.5-year-old children and in their parents, the numerical size of spherical anisometropia increased with myopia (p < 0.05 and p < 0.01, respectively). Cylindrical anisometropia did not exhibit such an increase. CONCLUSION: In Hong Kong children and their parents we found a tendency towards more spherical myopia in right eyes (laterality) in myopic cases. The numerical size of spherical anisometropia also increased in myopia in these groups. Cylindrical anisometropia appeared to be independent of spherical ametropia. In most cases right/left differences in both sphere and cylinder were small and our findings justify the use of data from one eye only in publications on refraction. PMID- 15043537 TI - Chemokine receptors and early activation markers in acute anterior uveitis. AB - PURPOSE: To study the peripheral expression of chemokine receptors and early activation markers in acute anterior uveitis (AU). METHODS: The expression of chemokine receptors CCR1, CCR2, CCR3, CCR5 and CXCR3, and activation markers CD25 (IL2R-alpha-chain), CD122 (IL2R-beta-chain) and CD69 were studied on peripheral blood mononuclear cells using flow cytometry in 10 patients with acute anterior uveitis and in 10 healthy controls. RESULTS: We found a highly significant up regulation of CCR5 (p < 0.001) on CD4+ T-cells in the blood of patients with AU compared to in healthy controls. CD69 was significantly higher on both CD4+ and CD8+ T-cells (p < 0.05 and p < 0.001, respectively). We also found a significantly higher expression of CD25 expressing CD4+ T-cells in patients with anterior uveitis (p < 0.05). This expression was directly correlated to the severity of disease (rho = 0.720, p < 0.05). CONCLUSION: Our data show that patients with acute anterior uveitis have a specific systemic immune activation. PMID- 15043538 TI - Time-lapse recordings of human corneal epithelial healing. AB - PURPOSE: The aim of this study was to design an experimental set-up for the study of human corneal epithelial wound healing in a controlled in vitro situation. METHODS: A time-lapse set-up was used. This allowed for pictures to be captured with a magnification ranging from x 80 to x 1800. Pictures were captured at 1-min intervals during the observation period, which lasted up to 4 days. Human corneal tissue was obtained from the Eye Bank or from surgery. A small, rounded lesion was produced in the corneal epithelium with a miniature drill. The specimens were placed in a mini-incubator; the camera focused on the epithelial lesion and continuously observed using the time-lapse set-up. RESULTS: The healing process of human corneal epithelium could be followed for several days. The initial healing response could be divided into a slow, a rapid and a consolidating phase. The first two phases lasted about 12 hours, and by then, epithelial cells covered the lesion. Depending on the origin of the tissue and the placement of the lesion, variations in the healing response could be seen. CONCLUSION: The time lapse technique makes it possible to study epithelial wound healing over time at the cellular level. Data collected in this way can fill the gap between in vivo studies, where, by nature, human wound healing studies are restricted, and cell culture techniques, where cellular responses in many cases differ from the in vivo situation. PMID- 15043539 TI - Evaluation of toxicity due to vital stains in isolated rat retinas. AB - PURPOSE: We investigated whether artificial aqueous humours are adequate incubation media compared with artificial cerebrospinal fluid (aCSF), and evaluated the retinal toxicity of two vital stains--trypan blue (TB) and indocyanine green (ICG)--and triamcinolone acetonide (TA) using isolated rat retinas incubated in artificial aqueous humours. METHODS: In experiment 1, retinal segments were isolated and incubated in aCSF, BSS plus, Opeguard Neo Kit, or phosphate-buffered saline (PBS). In experiment 2, retinal tissues were exposed to one of the agents and incubated in BSS plus. Retinal damage was assessed by morphological examination and biochemical assay, which measured lactate dehydrogenase (LDH) in the medium once every hour. RESULTS: In experiment 1, BSS plus was confirmed as a suitable incubation medium. In experiment 2, there were no significant changes in the retinas exposed to TB or TA. Tissues exposed to ICG showed damage in every retinal layer and significantly higher release of LDH. CONCLUSION: Exposure to ICG caused retinal damage in isolated rat retina tissue in our experimental model (in vitro). PMID- 15043540 TI - Efficacy of early systemic betamethasone or cyclosporin A after corneal alkali injury via inflammatory cytokine reduction. AB - PURPOSE: To evaluate the effects of early systemic administration of betamethasone or cyclosporin A (CsA) on inflammatory cytokine expression and corneal damage after alkali injury. METHODS: Sixty-five Wistar rat corneas injured with 1N NaOH were divided into three groups: untreated, betamethasone treated and CsA-treated. Both agents were administered systemically and daily during the first 7 days after injury. Interleukin (IL)-1alpha, -6 and -8 concentrations in the injured corneas were measured with ELISA at 2, 4 and 7 days after injury. Corneal damage was evaluated by scoring clinical findings. RESULTS: In untreated injured corneas, IL-1alpha, -6 and -8 were markedly elevated during the 7-day period following injury. Both betamethasone and CsA significantly suppressed IL-1alpha and IL-8 at day 4. Only betamethasone significantly suppressed IL-6 at days 4 and 7. Both agents significantly reduced corneal opacity at day 4. CONCLUSION: Early systemic administration of betamethasone or CsA after alkali injury may be of benefit by suppressing inflammatory cytokine expression in the cornea. PMID- 15043541 TI - Biglycan gene expression in UVR-exposed rabbit corneas. AB - PURPOSE: It is known that stromal proteoglycans play an important role in the hydration and transparency of the mammalian cornea. Proteoglycans have been described as a pathological deposit in climatic proteoglycan stromal keratopathy, which is associated with chronic ultraviolet radiation (UVR) exposure. The expression of dermatan sulfate proteoglycan biglycan in the cornea was thus studied after exposure of rabbit eyes to UVR. METHODS: New Zealand albino rabbit corneas were exposed to UVR at 310 nm at the dose producing biomicroscopically significant keratitis (0.47 J/cm2). Animals were killed 3, 7 and 28 days after exposure (five rabbits in each group). Five rabbits were used as controls and did not receive any UVR treatment. Expression of biglycan mRNA in the corneas was investigated by competitive reverse transcription-polymerase chain reaction (RT PCR). RESULTS: There was no expression of biglycan mRNA in the control group. In the UVR-exposed groups, biglycan mRNA had still not been expressed 3 days after exposure. The expression of biglycan mRNA was observed in all UVR-treated corneas 7 days after exposure (p < 0.05). By 28 days after UVR exposure the expression of biglycan mRNA had decreased (not statistically significant). CONCLUSIONS: There is no detectable biglycan gene expression in the normal rabbit cornea. Ultraviolet radiation exposure leads to a distinct expression of biglycan mRNA in the rabbit cornea that decreases 4 weeks after exposure, indicating the involvement of biglycan in the corneal repair process. Biglycan appears to be a novel marker of corneal wound healing. PMID- 15043542 TI - Transpupillary thermotherapy for idiopathic subfoveal choroidal neovascularization. AB - PURPOSE: To assess the efficacy of transpupillary thermotherapy (TTT) for the management of idiopathic subfoveal choroidal neovascularization (CNV). METHODS: This was a prospective, non-controlled, clinical case trial involving 21 eyes of 21 patients with a mean age of 36 years. All 21 eyes had idiopathic subfoveal CNV and were subjected to diode laser TTT. Laser beam size ranged from 1.2 mm to 3.0 mm and power settings ranged between 300 mW and 600 mW. The end-point was an area of no visible colour change. Pre- and post-laser evaluations of ETDRS visual acuity, contrast sensitivity, slit-lamp biomicroscopy and fluorescein angiography were carried out. RESULTS: The nine predominantly occult and 12 predominantly classic subfoveal membranes were followed-up for a mean of 5.1 months after TTT. Overall, 17 eyes (81%) showed improvement or stabilization in ETDRS visual acuity. Contrast sensitivity improved in 17 eyes (81%). Reading speed improved in 15 eyes (71%). Retreatment was needed in three eyes (14%). CONCLUSION: Transpupillary thermotherapy treatment is a potential strategy for treating idiopathic subfoveal CNV in both its classic and occult forms. PMID- 15043543 TI - A prospective longterm study of primary chronic angle closure glaucoma. AB - PURPOSE: To prospectively evaluate the longterm outcome of therapy for chronic primary angle closure glaucoma (PACG) and to assess the efficacy of medical and surgical treatment in terms of intraocular pressure (IOP) and visual field stabilization. METHODS: Seventy consecutive patients with chronic PACG, whose IOP remained > 21 mmHg despite a patent iridotomy, had their IOP controlled by medications or trabeculectomy performed without antimetabolites. They were followed over a 6-year period. Best corrected visual acuity, IOP (mean of annual diurnal variation readings), cup : disc ratio and visual fields were recorded. A trabeculectomy was performed if the IOP was not adequately controlled on maximal tolerable medical therapy or if there was a progression of the glaucomatous defect. Data from one eye of each patient were analysed; if both eyes met the inclusion criteria, one was randomly selected for the analysis. The baseline parameters were compared with those at the end of 6 years. RESULTS: A total of 46 eyes (65%) were controlled medically throughout the 6-year follow-up period, while 24 eyes (35%) required surgery. The mean IOP was 25.4 +/- 4.9 mmHg at baseline and 15.6 +/- 4.6 mmHg at 6 years follow-up (p < 0.001). Stereoscopic evaluation of the cup : disc ratio did not show a significant change from a mean of 0.6 +/- 0.18 at baseline to a mean of 0.64 +/- 0.2 at 6 years (p = 0.12). Progression of visual field defects was seen in seven eyes (10%), which had statistically larger cup : disc ratios (p = 0.04) and more extensive visual field deficits at the initial assessment (p = 0.04), and which also maintained higher levels of IOP (p = 0.03) over the 6 years of follow-up. CONCLUSIONS: Stable visual fields and good longterm IOP control were seen in 90% of chronic primary angle closure glaucoma eyes on medical/surgical therapy over 6 years. PMID- 15043544 TI - Macular haemorrhage following implantation of a posterior chamber collamer lens. PMID- 15043545 TI - A retinal pseudotear: optical coherence tomography view through retino-vitreal proliferations. PMID- 15043546 TI - Optical coherence tomography in the diagnosis of juvenile X-linked retinoschisis. AB - PURPOSE: To describe the value of optical coherence tomography (OCT) as a diagnostic tool in the diagnosis of X-linked retinoschisis. METHODS: We report three boys aged between 8 and 17 years, diagnosed with X-linked retinoschisis. During investigations they were examined with OCT (Zeiss Humphrey OCT 1, upgraded version). Single scans of the central posterior pole and the region around the vascular arcades were obtained. Two of the boys underwent full-field ERG according to ISCEV standards. Genetic analysis was performed in all three boys, with sequencing of the XLRS gene. RESULTS: The OCT results revealed a pattern with a cleavage of the retina in two distinct planes, one deep (outer retina) and one superficial. This was very obvious in one patient and a similar but not as pronounced pattern was seen in the other two cases. The two layers were superficially connected with thin-walled, vertical palisades, separated by low reflective, cystoid spaces, confluent and most prominent in the foveal region. CONCLUSION: Full-field ERG and/or DNA analysis are well known methods used for diagnosis of X-linked juvenile retinoschisis. In this paper, we suggest that OCT can also be a helpful diagnostic tool. PMID- 15043547 TI - Primary corneal myxoma. AB - PURPOSE: To present the clinical and histopathological characteristics of a corneal myxoma. METHODS: A 36-year-old, white woman presented with blurred vision and a whitish opacity on the left cornea. Clinical examination revealed a whitish, non-tender, 5 x 9 mm superficial tumour on the cornea. The tumour was excised and examined microscopically. RESULTS: The tumour was composed of spindle shaped cells in a myxomatous ground substance. Immunohistochemically, the tumour cells were positive for vimentin, muscle-specific antigen and smooth muscle antigen. The tumour was classified as a myxoma of the cornea. The origin of the tumour is discussed. CONCLUSION: This is the second reported case of a corneal myxoma without prior corneal disease. PMID- 15043548 TI - Are cataract and iris atrophy toxic complications of medical treatment of acanthamoeba keratitis? AB - PURPOSE: Two cases of acanthamoeba keratitis with almost identical clinical courses developed mature cataract and iris atrophy after prolonged treatment with chlorhexidine 0.02% and propamidine isethionate 0.1%. It is suggested that these complications were caused by the chemicals, rather than resulting from the amoebae-induced inflammation. METHODS: Penetrating keratoplasty was performed in both cases when perforation from peripheral ulceration was threatening. RESULTS AND CONCLUSIONS: The immediate results were good. From these two cases it seems possible that there is a maximum level of intensity of the medical treatment and also that surgery should be considered at an earlier stage, before intraocular complications develop. PMID- 15043549 TI - Fungal endophthalmitis caused by Paecilomyces variotii following cataract surgery: a presumed operating room air-conditioning system contamination. AB - PURPOSE: To report a case of delayed fungal endophthalmitis by Paecilomyces variotii following uncomplicated cataract surgery. To our knowledge this is the first reported case of postoperative endophthalmitis by this species. METHODS: We report the longterm clinical follow-up of an 83-year-old female who underwent uncomplicated sutureless, small-incision cataract surgery. She developed recurring uveitis 4 months after surgery. Vitreous tap and finally complete vitrectomy with removal of the capsular bag including the intraocular lens were performed. Fungi were studied by histopathology and culture. RESULTS: At histopathological examination, the fungi were found to be closely related with the capsular bag. A few mononuclear inflammatory cells were encountered. At culture, Paecilomyces variotii, a common ubiquitous non-pathogenic saprophyte, was identified. Despite systemic, intravitreal and topical antifungal therapy after vitrectomy the uveitis recurred several times, but no fungal organisms were isolated from the repeat intraocular specimen. At 18 months postoperatively the subject's visual acuity was finger counting at 2 metres. At the time of surgery the operating room air-conditioning system was undergoing repairs. Cases of fungal endophthalmitis after contamination from air-conditioning ventilation systems have been reported before, but none of the cases reported have been caused by P. variotii. CONCLUSION: P. variotii, a non-pathogenic environmental saprophyte, may be disastrous if introduced into the eye. International recommendations on the environmental control of the operating room air conditioning ventilation system should be strictly followed. No intraoperative surgery should be undertaken while the air-conditioning system is undergoing repairs or service. PMID- 15043551 TI - Ophthalmodynamometric measurement of orbital tissue pressure in thyroid associated orbitopathy. PMID- 15043550 TI - Laser photocoagulation for choroidal neovascular membrane associated with optic disc drusen. AB - PURPOSE: To report two cases of choroidal neovascular membrane associated with optic disc drusen which were successfully treated using argon laser photocoagulation. METHODS: Choroidal neovascular complications of optic disc drusen were diagnosed in two of our patients, a 36-year-old woman and a 14-year old girl. In both cases visual acuity was 20/100 in the affected eye. Fundus examination and angiography revealed a choroidal neovascular membrane. Computer tomography identified bilateral calcified drusen of the optic discs. Photocoagulation of neovascular tissues was immediately performed. RESULTS: Visual acuity improved progressively, reaching 20/20 10 months after treatment in one case and 20/30 in the other. No recurrence was observed during follow-up. CONCLUSION: Early diagnosis and immediate laser photocoagulation of vision threatening choroidal neovascular membranes associated with optic disc drusen helps stop progression towards the macula and improves longterm visual acuity. PMID- 15043552 TI - A guide to making a natural eyelid margin curvature in blepharoptosis surgery. PMID- 15043553 TI - A case of orbital lymphangioma showing considerable shrinkage after limiting dissection. PMID- 15043554 TI - Age-related macular degeneration is an inflammatory disease possibly treatable with minocycline. PMID- 15043555 TI - Vitreous haemorrhage in a 19-year-old Japanese woman using an oral contraceptive. PMID- 15043556 TI - Peripheral corneal thinning and silent corneal perforation in an HIV seropositive case. PMID- 15043559 TI - Viral attacks on the blood supply: the impact of severe acute respiratory syndrome in Beijing. PMID- 15043560 TI - NAT screening of blood donors for severe acute respiratory syndrome coronavirus can potentially prevent transfusion associated transmissions. AB - BACKGROUND: The severe acute respiratory syndrome (SARS) was first described in February 2003. Close contact with symptomatic patients appears to be the main route of transmission, whereas blood transfusion transmission could not be ruled out. STUDY DESIGN AND METHODS: A SARS coronavirus (SARS-CoV) detection kit developed by C. Drosten (Bernhard Nocht Institute, BNI) was used to amplify SARS CoV sequences from blood donor samples. We tested 31,151 blood donor samples in minipools of up to 96 samples. To validate the sensitivity of the assay, routine donor minipools (88 +/- 8 samples per pool) were spiked with plasma of an imported case of SARS or of a subsequently infected contact person, respectively. Gamma-irradiated cell culture supernatants of Vero E6 cells, infected with SARS CoV, were used as positive controls. RESULTS: None of 31,151 blood donors were positive for the presence of SARS. Two 96-member plasma pools that were each spiked with 100 microL of plasma of the German index patient or his wife, respectively, were positive. Overall, 0.85 percent of test results had to be considered invalid owing to negative internal controls. CONCLUSION: A real-time CoV PCR test is able to detect SARS-CoV in viremic blood donor samples even in the beginning of the disease when patients present minor clinical symptoms. Thus the assay could potentially help to prevent transfusion-associated SARS-CoV transmissions. PMID- 15043561 TI - Development of a sensitive PCR inhibition method to demonstrate HBV nucleic acid inactivation. AB - BACKGROUND: The evaluation of pathogen reduction technologies with relevant viruses currently contaminating the blood supply is limited by the availability of high-titer virus inocula and sensitive in vitro or in vivo infectivity assays. Because HBV infectivity can only be assessed by in vivo studies with chimpanzees, a sensitive PCR inhibition assay was developed to measure PEN110 inactivation of HBV. STUDY DESIGN AND METHODS: PCR amplification of 1.1 kb of HBV genome was optimized to determine DNA damage introduced by treatment with PEN110 in RBCs. Inactivation of duck HBV (DHBV) in RBCs, with measurement of the in vitro infectivity, was performed to validate the PCR assay. RESULTS: The PCR was highly specific and sensitive for amplification of the HBV genome and used to demonstrate a reduction of at least 7.2 and 8.1 log geq per mL within the first 18 hours of PEN110 treatment. PEN110 inactivation of DHBV was also achieved within the first 18 hours with a reduction factor of at least 5.0 log tissue culture infectious dose 50 percent per mL, suggesting that PCR inhibition is an alternative to infectivity assays. CONCLUSION: This study establishes PCR inhibition as a reasonable approach to assess the efficiency of PEN110 inactivation of human pathogens with human plasma donations that have been found to contain high titers of relevant agents during different stages of infection. PMID- 15043562 TI - Hb content-based transfusion policy successfully reduces the number of RBC units transfused. AB - BACKGROUND: RBC transfusions should be based on Hb content of the products instead of units. As high as a 50-percent difference in Hb content can be encountered between two units. In this study, 500 mL of whole blood (WB) generated RBCs according to their total Hb content was used to decrease the number of units ordered by using a new software called Hemosoft. STUDY DESIGN AND METHODS: Fifty-one consecutive patients were enrolled for the study. The median age and male-to-female ratio were 38 (range, 16-69) and 28:23, respectively. Patients' diagnoses were 38 acute leukemia, 11 lymphoma, and 2 multiple myeloma. When placing orders for RBCs, clinicians were asked to send the information about patient's actual body weight (ABW) and their actual and target Hb. The total blood volume of the recipient was calculated, and the aim was to find the best RBCs from the inventory to decrease two-unit orders to one unit, or four units to two or one unit, all done using Hemosoft. Posttransfusion Hb was checked 2 hours after transfusion. ABW and the median pretransfusion Hb level were 64 kg (range, 40-100) and 7.3 g per dL (5.6-9.8), respectively. RESULTS: A total of 104 RBC units were ordered for 51 patients. In 49 (96.1%), the order was for two units and in 2 (3.9%) was for three units. Hemosoft successfully found a suitable match in 62.7 percent (32/51) of the orders. Actual number of RBCs transfused was two units (41.25) and one unit (58.8%), respectively. In total, 72 units were transfused with a reduction of 30 percent (72/104) from the original order. The median target Hb level was 9.3 g per dL (range, 7.0-11.3) and overall success rate to achieve it was 96.6 percent (range, 73.8-124.2). There was no impact of inventory size, but the ABW of the recipient was found significant to have a successful match unit (58.8 +/- 11.0 kg vs. 75.0 +/- 8.4 kg, respectively; p=0.0001). CONCLUSION: We clearly demonstrated that the number of RBC transfusions could be minimized by the rational use of the Hb content of the units. Although the total numbers of patients who shall benefit from this approach seems to be limited, it allows us to use high-Hb-content units rationally by using an in-house blood-banking software. PMID- 15043563 TI - The effect of universal leukoreduction on postoperative infections and length of hospital stay in elective orthopedic and cardiac surgery. AB - BACKGROUND: A before and after study was undertaken to investigate the effect of universal leukoreduction (ULR) in the UK on postoperative length of hospital stay (LOS) and infections. STUDY DESIGN AND METHODS: Consecutive patients undergoing elective coronary artery bypass grafting or total hip and/or knee replacement in 11 hospitals received non-WBC-reduced RBCs before implementation of ULR (T1, n=997) or WBC-reduced RBCs after implementation of ULR (T2, n=1098). RESULTS: Patients in T1 and T2 were comparable except patients in T2 received on average more units of RBCs but had lower discharge Hct levels. Postoperative LOS (T1, 10 +/- 8.9 days; T2, 9.6 +/- 6.9 days) and the proportion of patients with suspected and proven postoperative infections (T1, 21.0%; T2, 20.0%) were unchanged before and after ULR (LOS, hazard ratio 1.01, 95% CI 0.92-1.10; infections, OR 0.83, 95% CI 0.77-1.02). Subgroup analysis showed no significant interaction between storage age or dose of blood on responsiveness of primary outcomes to ULR. Secondary outcomes were unchanged overall. Analysis by surgical procedure gave conflicting results with both increased mortality (p=0.031) and an increased proportion of cardiac patients with proven infections (p=0.004), whereas the proportion of orthopedic patients with proven infections was reduced (p=0.002) after ULR. CONCLUSION: Implementation of ULR had no major impact on postoperative infection or LOS in patients undergoing elective surgical procedures who received transfusion(s). Smaller effects, either detrimental or beneficial of ULR, cannot be excluded. PMID- 15043564 TI - Decreased RBCTs after reduced intensity conditioning allogeneic stem cell transplantation: predictive value of prior Hb level. AB - BACKGROUND: RBCT (RBCT) requirements of stem cell transplant (SCT) recipients are often substantial and may be related to transplant type. STUDY DESIGN AND METHODS: An analysis was done of RBCT requirements and Hb recovery kinetic in the first 60 days after HLA-identical sibling allogeneic SCT in a series of 110 consecutive patients treated for various malignant diagnoses. Patients were prepared with either an antithymocyte globulin (ATG) and reduced intensity chemotherapy-based conditioning (RIC) (n=64) or a myeloablative conditioning regimens (MAC; n=46). Patients received marrow (n=64) or PBPCs (n=46). RESULTS: Overall, intensity of conditioning regimen (RIC vs. MAC; p=0.0005) and graft source (PBPC vs. marrow; p<0.0001) independently predicted RBCT requirements. Hb recovery was accelerated after RIC when compared to MAC allo-SCT (p=0.02). In RIC patients, RBCTs were inversely correlated to Hb level before conditioning (p<0.0001) and the dose of ATG (p=0.009). Moreover, Hb level before allo-SCT significantly influenced Hb recovery kinetic after RIC but had no impact on RBCT requirements and Hb recovery after MAC. CONCLUSION: Thus, RIC conditioning creates a different pattern of erythropoiesis recovery as compared to a MAC regimen and suggest a need for studies aimed at further reducing RBCT and accelerating Hb recovery. PMID- 15043565 TI - A new strategy for the prevention of IgA anaphylactic transfusion reactions. AB - BACKGROUND: Management of patients with clinically significant anti-IgA is difficult and unsatisfactory in many aspects. PATIENTS AND METHOD: A 40-year-old man with common variable immunodeficiency had a previous history of anaphylaxis after an intramuscular immunoglobulin administration. His serum contained anti IgA, and he required immunoglobulins for recurrent infections. RESULTS: The administration of intravenous immunoglobulins (IVIgG) containing less than 0.1 mg per mL IgA led to an anaphylactic reaction after the transfusion of only 2 to 3 mL. The same IVIgG charge was subsequently pretreated with freshly separated autologous plasma and given to the patient on three consecutive days without any reaction (1.25, 10, and 10 g each in 400 mL plasma). Anti-IgA activity did not increase, and the patient was treated again without complications. DISCUSSION: Ex vivo pretreatment of IVIgG preparations with autologous plasma appears to be safe and useful in the management of patients with clinically significant anti-IgA. To achieve a significant IgA blockage, the preparation to be used should not contain large amounts of IgA. CONCLUSION: The strategy described here appears to be safe and may help prevent anaphylaxis in many instances. PMID- 15043566 TI - Postdelivery levels of anti-D IgG prophylaxis in D- mothers depend on maternal body weight. AB - BACKGROUND: Current recommendations for anti-D prophylaxis for women who deliver a D+ offspring vary from country to country, and the introduction of new reagents require pharmacokinetic studies that show serum levels after the injection. Serum levels of anti-D may depend on the maternal body mass index (BMI). STUDY DESIGN AND METHODS: Serum concentrations of total anti-D IgG and IgG1-4 subclasses were determined by flow cytometry in 26 D- women, who had received prophylaxis after delivery of a D+ offspring. Blood samples were drawn on Days 1, 2, 3, and 14 after injection, and the BMI was recorded. RESULTS: Anti-D levels increased continuously in all women during the first 3 days. The increase was significantly affected by the BMI if higher than 27 kg per m2 (p<0.001). The higher the BMI, the less was the increase of serum anti-D. Mean peak levels 72 hours after injection was 89 ng per mL in lean women, but estimated levels were 28 to 60 percent lower in women with a BMI of 28 to 40 kg per m2. The effect of a BMI higher than 27 kg per m2 on anti-D was not gradual but progressive. Similarly, the BMI affected serum concentrations of anti-D subclasses IgG1-4 (p<0.001). CONCLUSION: The BMI needs consideration for the adjustment of the dosage of anti D, provided its bioavailability to suppress alloimmunization is reflected by measurable amounts in the serum. PMID- 15043567 TI - Liquid storage, shipment, and cryopreservation of cord blood. AB - BACKGROUND: Cord blood banking requires methods for shipping and storage. This study examines the influence of shipping via overnight courier on postthaw viability of cord blood. STUDY DESIGNS AND METHODS: Anticoagulated cord blood was divided with one sample diluted 1:1 using STM-sav (a storage solution) and the other undiluted. Units were shipped from Minneapolis to Memphis and returned, RBC depleted, cryopreserved, stored for 14 days, and thawed. MNC counts, percent viable cells, quantity of CD34+ cells, and frequency of CFU-GM were measured. Temperature during shipment was continuously monitored. RESULTS: Preliminary studies showed the packing and processing protocol influenced the temperatures experienced during shipping. Samples achieved temperatures below 10 degrees C within 4 to 8 hours with a few units dropping near or below 1 degrees C with cold ambient temperatures. The MNC recovery, CD34+45+ recovery, and frequency of CFU GM for samples that were shipped were comparable to those observed using static liquid storage. The postthaw viable cell recovery was greatest for storage and shipping times of 24 hours and decreased when the storage and shipping times were longer. CONCLUSION: Ambient conditions and the packing and processing protocol influence the temperature history of the sample. Samples stored beyond 24 hours in liquid storage and shipping exhibit a decreased postthaw recovery. PMID- 15043568 TI - Peripheral blood circulating immature cell counts predict CD34+ cell yields in G CSF-induced PBPC mobilization in healthy donors. AB - BACKGROUND: It has been previously reported that the number of circulating immature cells (CIC) in peripheral blood (PB) estimates the number of CD34+ cells collected in G-CSF plus chemotherapy-induced PBPC mobilization. The correlation of CIC counts in PB with CD34+ cell yield and its usefulness was evaluated in G CSF-induced PBPC mobilization for healthy donors. STUDY DESIGN AND METHODS: CIC counts in PB and CD34+ cell counts in the apheresis product from 122 collections were assessed, and the relationship between these two variables was evaluated with the Pearson rank correlation analysis, the chi-squared test, and the U-test. RESULTS: CIC counts were correlated weakly with the number of CD34+ cells per L of blood processed in the apheresis product (Pearson rank correlation analysis; r=0.357, p<0.0001). When a level of 1.7 x 10(9) CICs per L was selected as a cutoff value, the sensitivity and specificity for collecting more than 20 x 10(6) CD34+ cells per L of blood processed were 63.6 and 77.5 percent, respectively. CONCLUSION: The present study suggests that the number of CICs in PB may estimate the number of CD34+ cells collected. The data indicate that CIC counts above 1.7 x 10(9) per L can be used as a good predictor for PBPC collections containing more than 20 x 10(6) CD34+ cells per L of blood processed in a single apheresis procedure. PMID- 15043570 TI - Cellular therapy services provided by blood centers and hospitals in the United States, 1999: an analysis from the Nationwide Blood Collection and Utilization Survey. AB - BACKGROUND: The 2000 Nationwide Blood Collection and Utilization Survey was designed to assess cellular therapy product services in US blood centers and hospitals. STUDY DESIGN AND METHODS: Questionnaires were returned by 2,040 institutions. Data were analyzed for 30 quantitative variables related to cellular therapy product activities. RESULTS: 269 institutions, including 231 (12.2%) of the hospitals, 37 (25.9%) of the blood centers, and one cryobank, performed HPC services. Collected PBSC (20,517) and cord blood products (12,628) far exceeded bone marrow (1,572), lymphocytes (578), and cultured cells (344). PBPC collections dropped 36.5 percent since the 1997 survey. Cord blood accounted for 35.4 percent of collections and 39.5 percent of products processed, but only 1.9 percent of infusions. CONCLUSIONS: Most cellular therapy services in hospitals and blood centers were HPC-related. The dramatic drop in PBPC collections since 1997 reflects the decline in autologous PBPC transplantation for breast cancer. Cord blood's high collection-to-infusion ratio demonstrates a substantial resource expenditure for banking a product for future clinical needs. Lymphocytes and cultured cell products contributed minimally to activities in this survey, but will likely increase in the future. Data from additional academic and commercial manufacturers of cellular therapy products should be included in future surveys. PMID- 15043569 TI - Low-dose lenograstim to enhance engraftment after autologous stem cell transplantation: a prospective randomized evaluation of two different fixed doses. AB - BACKGROUND: G-CSF is used to enhance hematopoietic recovery after autologous stem cell transplantation (ASCT), but the optimal dose of G-CSF during engraftment has not been established. The medical cost of ASCT is a serious financial burden in developing countries, and G-CSF is the most costly drug used in this procedure. We evaluated whether a lower, vial-size fitted dose of lenograstim is clinically equivalent to a higher fixed dose. STUDY DESIGN AND METHODS: A prospective randomized study was performed on 33 patients (11 non-Hodgkin's lymphoma, 8 multiple myeloma, 14 breast cancer) undergoing ASCT. Patients were randomly administered 100 micro g or 250 micro g lenograstim daily starting on the next day of ASCT, with a minimum infusion of 3 x 10(6) CD34+ cells per kg. RESULTS: For both lenograstim doses, median time to neutrophil engraftment was 9 days and median time to PLT engraftment was 11 days. Episodes of clinically documented infections were 10 per 379 patient-days in the 100 microg per day group and 10 per 320 patient-days in the 250 microg per day group. There were no between-group differences in requirements for transfusion of RBCs or PLTs. Duration of hospitalization was 16 days for the 100 microg per day group and 17 days for the 250 microg per day group. Daily lenograstim dose per patient's body weight and total amount of lenograstim used during ASCT were both significantly lower in the 100 microg per day group. CONCLUSION: Administration of 100 microg per day of lenograstim showed comparable clinical efficacy to 250 microg per day lenograstim for immediate hematopoietic recovery after ASCT. Use of the lower dose was associated with lower overall lenograstim usage and lower cost. PMID- 15043571 TI - Viable CD34+ stem cell content of a cord blood graft: which measurement performed before transplantation is most representative? AB - BACKGROUND: Patient survival in allogeneic cord blood transplantation is critically dependent on total nucleated cell (TNC) count or total CD34+ cell count per kg of body weight. Theoretically, viable CD34+ cell measurement at the time of infusion should give a better indication of the suitability of a certain transplant. The relation between measurements on different samples and viable CD34+ cell count on the graft itself was analyzed. STUDY DESIGN AND METHODS: Viable CD34+ cells were measured with a no-wash, single-platform technique with 7 aminoactinomycin D. Analysis was performed before freezing on the cord blood, after freezing and thawing on the cord blood unit itself, and on various samples. RESULTS: Cord blood volume correlated poorly with viable CD34+ cell content (r=0.24) as did initial TNC count and WBC count (r=0.57 and r=0.48, respectively). In contrast, viable CD34 cell content determined before freezing correlated well with viable CD34 cell content of the graft (r=0.91) but was on average 25 percent higher than after freezing and thawing. The best correlations with the CD34+ cell content of the cord blood unit were obtained with CD34 cell measurements on a separate cryovial (r=0.95). These CD34 cell measurements on frozen samples were found to be very reproducible (r=0.96). CONCLUSION: Viable CD34 cell count of the graft is both accurate and precise when measured on a separate sample frozen together with the cord blood unit. This measurement can be performed by the transplant center to exclude between-laboratory variability. PMID- 15043572 TI - A preclinical xenotransplantation animal model to assess human hematopoietic stem cell engraftment. AB - BACKGROUND: Delayed megakaryocytic engraftment occurs in approximately 8 percent of patients undergoing autologous transplantation with PBPCs, and a reliable assay to predict engraftment is not yet available. STUDY DESIGN AND METHODS: The correlation between human cell engraftment in a mouse xenotransplantation model with the rate of megakaryocytic recovery for individual patients after autologous PBPC transplantation was evaluated. Engraftment into nonobese diabetic (NOD) severe combined immunodeficient (SCID) and NOD-SCID-beta2m null mice was compared for patients with rapid (11 days) PLT recovery (good engrafters, GEs) versus those with delayed (18 days) PLT engraftment (poor engrafters, PEs). PBPCs (1 x 10(6) CD34+ cells) were transplanted into sublethally irradiated (300 cGy) mice, and human WBC and human PLT engraftment were analyzed by FACS in the blood weekly. Human WBCs and human CFU-megakaryocytes (Mks) in the marrow were determined 6 to 7 weeks after transplant. RESULTS: Six PEs and five GEs were analyzed. Four of six PEs showed no human cell engraftment, whereas five of five GEs showed multilineage human hematopoiesis including the presence of CFU-Mks. Human WBC engraftment and human CFU-Mks differed significantly between GEs and PEs (p<0.01). NOD-SCID-beta2m null had significantly higher levels of human engraftment than NOD-SCID mice (p<0.05). The two PEs whose PBPCs were capable of engrafting in the mice had underlying liver abnormalities that may have played a role in their delayed engraftment. CONCLUSIONS: Time to PLT recovery in patients correlates strongly with human PLT and human WBC engraftment and with the number of human CFU-Mks (p<0.05) in a xenogeneic transplant model. This model may be useful for future studies to test therapeutic strategies for enhancement of engraftment. PMID- 15043573 TI - Recombinant glycophorins C and D as tools for studying Gerbich blood group antigens. AB - BACKGROUND: The Gerbich blood group system antigens are carried on glycophorin C (GPC) and glycophorin D (GPD) and variants thereof. These glycoproteins have been expressed in a heterologous system to study the individual antigens and to determine whether Ana is antithetical to Ge2. STUDY DESIGN AND METHODS: cDNAs encoding GPC, GPD, GPC.Yus, GPC.Ge, GPC.Lsa, and GPD.Lsa were transfected and stably expressed in a human embryonic kidney cell line (293T). Individual Gerbich antigens were analyzed with MoAbs and human polyclonal antibodies by flow cytometry and immunoblotting. Recombinant GPD and GPD.Ana were expressed transiently and analyzed for expression of Ge2 and Ana antigens. RESULTS: All recombinant variants were detected with sialidase-resistant and -sensitive anti Ge2, anti-Ge3, and anti-Ge4. Ge4 antigen expression was depressed in GPC.Ls(a) transfectants as well as on Ls(a+) RBCs. GPD.An(a) recombinant protein expressed Ana and Ge2 antigens. CONCLUSION: Cell lines stably expressing glycosylated Gerbich proteins were developed in a heterologous system by transfecting individual variant forms of GPC and GPD. Unexpectedly, it was found that Ge4 antigen is reduced in both the GPC.Ls(a) recombinant and the Ls(a+) RBCs. It was also shown that Ana and Ge2 antigens were expressed on a single GPD.An(a) protein and, therefore, they cannot be antithetical. PMID- 15043574 TI - Molecular and serologic characterization of DWI, a novel "high-grade" partial D. AB - BACKGROUND: Accurate D antigen identification is essential for pretransfusion and prenatal evaluation to prevent anti-D alloimmunization. Quantitative and qualitative D variants may pose typing problems and require particular consideration because of differing potential for anti-D induction. STUDY DESIGN AND METHODS: A novel partial D, DWI, was discovered in an anti-D-alloimmunized D+ Austrian woman. This D variant was investigated by RHD genotyping and nucleotide sequencing, as well as characterization of its serologic properties. RESULTS: The proposita exhibited a single-nucleotide exchange in RHD Exon 7 (1073T>C) predicting a Met358Thr substitution in the sixth extracellular loop of the RhD polypeptide. All DWI individuals identified (the proposita and two relatives) were genotyped DWIdCcee, which, together with the family tree, was highly suggestive of a DWICe haplotype association. Epitope mapping studies revealed only minor D antigen modification with weakening but not loss of epitopes D1.1, D9.1, and D16.1. Antigen density varied individually between 8000 and 8600 D sites per erythrocyte. No known low-frequency Rh antigen was detected. Despite the highly retained D epitope composition, the DWI proposita's serum sample contained alloanti-D from an immunization event many years earlier. CONCLUSION: The findings of this investigation emphasize the possible clinical significance of "high-grade" partial D variants that are likely to be missed by routine serology. PMID- 15043575 TI - Significance of human neutrophil antigen-2a (NB1) expression and neutrophil number in pregnancy. AB - BACKGROUND: Expression of human neutrophil antigen-2a (HNA-2a) is greater in women than in men. The size of the HNA-2a-positive neutrophil population increases with pregnancy. STUDY DESIGN AND METHODS: T he relationship between HNA 2a expression on neutrophils and monocytes, and their relative numbers, was investigated. HNA-2a expression shown as the size of the HNA-2a-positive cell population and the fluorescence intensity on the cells was analyzed using flow cytometry. This investigation was done among 165 pregnant women during pregnancy and postpartum. RESULTS: In normal pregnancy, numbers of neutrophils and monocytes changed in relation to HNA-2a expression. HNA-2a was also expressed intensively on monocytes from some pregnant women in the first and second trimesters. HNA-2a expression and the number of cells markedly decreased postpartum. In threatened premature labor, the number of neutrophils decreased earlier than in normal pregnancy. CONCLUSION: HNA-2a expression may increase soon after fertilization. In this study, the results indicate that the change in the number of neutrophils and monocytes is related to HNA-2a expression. PMID- 15043577 TI - Adjusting our management of female blood donors: the key to an adequate blood supply. PMID- 15043576 TI - Upper-extremity deep venous thrombosis complicating whole-blood donation. AB - BACKGROUND: Up to 36 percent of blood donors may experience a donation-related complication. Fatigue, bruises, hematomas, and vasovagal reactions comprise the great majority of donor reactions and injuries. Serious complications are rare. CASE REPORT: A 20-year-old female taking the third-generation oral contraceptive desogestrel/ethinyl estradiol and ethinyl estradiol (Mircette) developed bruising and increased pain and swelling of her right arm over a 5-day period after whole blood donation. She was a first-time donor and the venipuncture was reported as being mildly traumatic. There was no personal or family history of thrombosis. RESULTS: Ultrasound examination of her upper extremity revealed the presence of a deep venous thrombosis that required treatment with enoxaparin sodium for 5 days and warfarin for 6 months. Evaluation for thrombophilia was negative. The only risk factor for thrombosis was use of oral contraceptives. CONCLUSION: Although serious complications from whole-blood donation are rare, they may occur. Deep venous thrombosis should be considered in a donor presenting with increasing pain and swelling after blood donation. PMID- 15043578 TI - Recruiting blood donors: challenges and opportunities. PMID- 15043579 TI - Proteomics in transfusion medicine. PMID- 15043580 TI - Measurement of bleeding severity: a critical review. PMID- 15043581 TI - Utility of in vitro tests in predicting the in vivo viability of stored PLTs. PMID- 15043582 TI - Pre-transfusion phenotype matching for sickle cell disease patients. PMID- 15043583 TI - Detection of fat particles in postoperative salvaged blood in orthopedic surgery. PMID- 15043584 TI - Hb screening of blood donors. PMID- 15043585 TI - Non-progression of HIV infection 20 years after diagnosis. PMID- 15043586 TI - Red cell freezing and its impact on the supply chain. AB - Red blood cells (RBC) can be frozen in glycerol solutions and stored for many years. Thawed RBC must have the glycerol removed, but the recovered cells have normal survival in humans. Freezing has been used to store RBC of rare phenotypes for more than 40 years. In the 1960s and 1970s, when medical technology and blood use were expanding rapidly and liquid whole blood and RBC storage were limited to 3 weeks, many attempts were made to expand the use of frozen RBC for meeting the needs for a stable blood supply and to have RBC reserves for emergencies. These attempts have largely been abandoned because of the cost of freezing, storing and processing, better management of the larger and longer lived RBC inventory, concerns about the safety of stored RBC that have not received the most up-to date testing and the losses associated with the short shelf life of thawed RBC. New automated frozen RBC processing systems will potentially allow extending the outdate of thawed RBC to 2 weeks, but will not materially effect the costs or losses associated with the use of frozen RBC. RBC freezing will have little effect on the logistics of blood supply. PMID- 15043587 TI - What population factors influence the decision to donate blood? AB - Donations of blood in most of the developed world are by nonremunerated volunteers. As such factors, which impact upon the motivation of individuals to donate, are critical to achieving a stable supply, we analyze the factors, which influence the decision to donate. Using data on 130,356 [corrected] individuals from 1999 and 2000, we investigate which factors determine the proportion of blood donors in postcode areas. Variables analyzed include blood donation status and sociodemographic characteristics in order to provide information on donation decisions. We find the proportion of donors in an area is significantly increased by higher proportions of women aged 20-29 and 40-49, and of men aged 60-69. Conversely, a higher proportion of males aged 20-29 significantly reduces proportion of donors. Also, a higher proportion of individuals born overseas significantly reduces the proportion of blood donors in an area. To increase supplies, blood collection agencies should target specific groups. Young men and men and women aged 30-49 need to be encouraged to donate. Collection agencies also need to target individuals who are born overseas to participate in the process. Using these results to aid targeting should help to maintain the blood supply. PMID- 15043588 TI - Knowledge, attitudes, beliefs and motivations towards blood donations among blood donors in Lagos, Nigeria. AB - Summary. interviewed with questionnaires on their knowledge, attitudes, beliefs and motivations about blood donations. It was found that a large number of them (92.9%) donated because of the benefits they will obtain from the hospital. Such benefits include antenatal registration (67.1%) and saving the lives of relations (25.8%). Even though many of the donors are educated (98.9%), majority of whom have university degrees (36.1%) and have heard about blood donation before, 52.4% of them believe they can contact human immunodeficiency virus (HIV) and/or hepatitis infection from blood donation. A good number (47.0%) are afraid of what they regard as side effects, such as weight loss (23.8%), sexual failure (5.9%), high blood pressure (5.2%), sudden death (3.3%), and convulsion (1.47%). About 41.0% prefers certificates as an incentive for donation, whereas 13.6% prefers money; less than 3% will like their names announced or published on the media and 2.58% will donate for nothing. It is recommended that an intensive blood donation campaign should be maintained. This will allow people to be well informed, turning the positive attitude of saving life through blood donation to a regular practice. PMID- 15043589 TI - Evaluation of the future supply and demand for blood products in the United Kingdom National Health Service. AB - In addition to the clinical management of blood supplies, the UK National Blood Service (NBS) examines short-term supply and demand to predict any potential shortages in blood supplies. However, very little data are available on the medium- and long-term trends. This paper describes blood product use in a United Kingdom Hospital Trust in 1999 and combines it with donor information to project the effect of demographic changes on demand and supply over the next 25 years. Overall, 2801 (4%) inpatients received transfusions of blood products. The proportion of inpatients receiving blood products increased with age. Patients aged >70 years used 46% of the total blood product supply, whereas patients aged <30 years used 10%. The estimated total cost associated with blood product use was pound 2 million during 1999 (2001-2002 prices). The results show that within 20 years, demand for blood products is expected to increase by 20% relative to the supply. The estimates in this study are a valuable aid to better long-term planning of supplies. As there is a paucity of data concerning blood product use in UK hospitals and blood use varies greatly between hospitals, further studies are required to guide blood product use policy. PMID- 15043590 TI - Reducing red cell transfusion by audit, education and a new guideline in a large teaching hospital. AB - Safety concerns combined with the greatly increased costs and difficulties of maintaining the blood supply are major considerations for transfusion services. Previous local surveys demonstrated that hospital blood use at our hospital could be improved. Excessive cross-matching, unnecessary transfusion and high return rates of unused blood were commonplace. Transfusion practice was audited over a 3 month period. An education package with guidelines for transfusion was delivered to all clinician groups within the hospital, over the following 9 months. The audit was repeated exactly 1 year later at the same time period. During the second audit, inpatient hospital numbers increased by 1.02% (from n = 7262 to n = 7336) but no differences in length of stay, cardiovascular morbidity or mortality were demonstrated. Twenty percent (n = 254, 2002; n = 316, 2001) fewer patients received blood, and the number of red cell packs used reduced by 19% (from n = 1093 to n = 880). Total number of patients transfused reduced from 4.4% to 3.5% which, as an absolute difference, is a reduction of 0.9% (CI 0.3-1.5, P = 0.006). The audit, guideline and education package had a major impact on red cell use within the hospital with no adverse effects. Blood use can be improved by the implementation of a suitable education package and guideline. If it is possible to replicate the results of this education programme nationwide, the effect on blood use, with subsequent savings and enhanced patient safety could be significant. PMID- 15043591 TI - Feasibility of a restrictive red-cell transfusion policy for patients treated with intensive chemotherapy for acute myeloid leukaemia. AB - Red-cell transfusions are required for symptomatic treatment of severe anaemia caused by intensive chemotherapy. Concerns about the transfusion-related complications, such as infections (e.g. the very low risk of human immunodeficiency virus (HIV)/hepatitis C virus (HCV) transmission and the risk of postoperative infections), haemolytic transfusion reaction, immunological effects and the costs, prompt a reevaluation of the transfusion practice. Retrospective analysis of prospectively collected data on 84 patients with acute myeloid leukaemia (AML), who were treated with combination chemotherapy between June 1, 1997 and December 7, 2001, was performed. The use of red-cell transfusions with a restrictive transfusion policy (haemoglobin = 7.2-8.8 g dL(-1), dependent on age and symptoms, n = 38) was compared with a more liberal transfusion trigger (haemoglobin = 9.6 g dL(-1), n = 46). The number of units transfused was recorded. Signs and symptoms of anaemia, chemotherapy-related effects and complications were investigated for both transfusion policies. The more restrictive transfusion policy led to a significant decrease of 11% of red blood cell (RBC) transfusions in patients with AML. No significant differences were found in the incidence of infections, number of platelet units transfused, bleeding complications, cardiac symptoms or response to chemotherapy. The more restrictive transfusion policy was feasible in this clinical setting, and it might be concluded that a restrictive transfusion policy is safe in supporting clinical patients treated with intensive chemotherapy for AML. PMID- 15043592 TI - Comparison of von Willebrand factor antigen, von Willebrand factor-cleaving protease and protein S in blood components used for treatment of thrombotic thrombocytopenic purpura. AB - Replacement of normal levels of von Willebrand factor-cleaving protease (VWF:CP, ADAMTS13) activity from infused plasma is important in plasma exchange (PEX) for the treatment of thrombotic thrombocytopenic purpura (TTP) patients. We have studied the VWF:CP activity, VWF multimer distribution, VWF:Ag, protein S (PS) activity and free PS antigen levels in fresh frozen plasma (FFP), cryosupernatant (CSP) and virally inactivated components treated with methylene blue/light (MB) or solvent detergent (SD) processes. VWF:CP activity was normal in all components tested and was retained following overnight storage at room temperature. CSP and SD plasma contained reduced levels of the highest molecular weight VWF multimers. Protein S activity was reduced below the normal range in SD plasma, but within the normal range for the other components tested. Virally inactivated SD- and MB treated plasma may be an effective alternative to FFP and CSP in PEX for TTP. Reduced PS activity in SD plasma may predispose to venous thromboembolism, especially if infused in large volumes. PMID- 15043593 TI - Complement activation in prestorage leucocyte-filtered plasma. AB - Complement activation and generation of pro-inflammatory cytokines occur during storage of blood components. Prestorage leucocyte filtration of platelet concentrates and red cells diminishes the accumulation of leucocyte-derived cytokines during storage, however, transfusion reactions are not eliminated. We investigated inflammatory mediator release during storage of plasma and whole blood and the effect of prestorage leucocyte filtration of plasma. Twenty-four blood units were collected from healthy blood donors and stored for 35 days. Eight units were stored as whole blood, eight units as plasma and eight units as prestorage filtered plasma. Samples were collected weekly for analyses of potassium, leucocytes, free plasma haemoglobin, complement activation (C3a and SC5b-9) and pro-inflammatory cytokines [interleukin (IL)-6, IL-8 and tumor necrosis factor (TNF)-alpha]. Elevated levels of C3a and SC5b-9 were registered in filtered plasma, from the beginning of storage. C3a levels increased during storage. There was a higher rate of change during storage in C3a (P < 0.01) and SC5b-9 (P < 0.05) in plasma compared with filtered plasma. Interleukin (IL)-8 is released in whole blood. The cytokine levels generated in plasma and filtered plasma were low. Complement activation is present in whole blood, plasma and filtered plasma during storage. Prestorage filtration of plasma activates the complement cascade but does not influence cytokine generation. PMID- 15043594 TI - Tazobactam-induced haemolytic anaemia, possibly caused by non-immunological adsorption of IgG onto patient's red cells. AB - A patient with pneumonia was treated with Tazocin (piperacillin/tazobactam). However, the expected haemoglobin (Hb) increment after transfusion was not achieved. Plasma bilirubin and lactate dehydrogenase were raised. The direct antiglobulin test (DAT) was positive (4+) for immunoglobulin G (IgG) only, but no RBC antibodies were demonstrable in the plasma or an eluate from the patient's RBCs. Drug-induced haemolysis was suspected. After discontinuing Tazocin administration, Hb and bilirubin levels returned to expected values. The patient's plasma gave a positive (3+) indirect antiglobulin reaction only with RBCs pretreated with tazobactam. However, random patient plasmas also gave weak (+/- to 1+) reactions, indicating non-immunological adsorption of IgG onto RBCs rather than specific anti-tazobactam antibodies. Subsequently, plasma samples with varying IgG levels (0.8-89.7 g L(-1)) were tested against RBCs pretreated with tazobactam. The amount of plasma IgG non-immunologically adsorbed onto the drug-coated RBCs was found to correlate directly with the plasma IgG level. The patient had a high plasma IgG level (41.6 g L(-1)) which explains why the antiglobulin test was stronger with the patient's plasma than with random plasma samples. Previous reports (Garratty & Arndt, (1998) British Journal of Haematology, 100, 777-783; Arndt & Garratty (2000) Transfusion, 40, 29S) suggested that non-immunological coating of RBCs with IgG may affect RBC survival; our results would support that suggestion. This is the first reported case of haemolytic anaemia associated with tazobactam. PMID- 15043595 TI - Guidelines for compatibility procedures in blood transfusion laboratories. PMID- 15043596 TI - A two-phase audit of fresh frozen plasma: a regional approach. PMID- 15043597 TI - Estimation of HIV seroprevalence in blood bank camps in Kolkata, India. PMID- 15043598 TI - Nucleic acid testing for virus screening in Polish blood donors. PMID- 15043599 TI - Walking and motoring: fitness and the social organisation of movement. AB - This essay applies the approach of the relational model to the quotidian modes of contemporary movement, walking and motoring, in order to analyse fitness. The social model of disability provides a tool for de-individualising fitness discourse, as well as for embedding fitness within sociomaterial and cultural contexts. Fitness, or 'being able to', is seen as an outcome of the interpenetrations and interactions of mind-bodies moving within their environments. The analysis indicates that the social model has general relevance for the sociology of health and illness. Here, it is extended beyond the study of disability to encompass fitness. In future, the social model may prove fruitful in the analysis of other socially situated conditions as well. PMID- 15043600 TI - Demystifying disability: a review of the International Classification of Functioning, Disability and Health. AB - The paper describes and evaluates the theoretical underpinnings of the International Classification of Functioning, Disability and Health (ICF), and develops the proposition that its conceptual framework provides a coherent, if uneven, guide through the competing conceptions of disability. To date, however, there has been little evaluation of the theoretical efficacy of the ICF. In seeking to redress this, the paper develops the argument that the ICF fails to specify, in any detail, the content of some of its main claims about the nature of impairment and disability. This has the potential to limit its capacity to educate and influence users about the relational nature of disability. The paper develops the contention that three parts of the ICF require further conceptual clarification and development: (a) (re)defining the nature of impairment; (b) specifying the content of biopsychosocial theory; and (c) clarifying the meaning and implications of universalisation as a principle for guiding the development of disability policies. PMID- 15043601 TI - Women's experiences of cervical cellular changes: an unintentional transition from health to liminality? AB - Cervical cancer screening is a preventive intervention directed towards women to both detect cervical cancer and identify those at risk for developing this disease. It has been argued that participation in screening programmes and early detection situations may lead to new kinds of sickness experiences. This article is based on qualitative phenomenological hermeneutical analysis of interviews with women who have received abnormal Pap smear test results through a population based outreach screening programme in urban Sweden. The aim of this article is to illuminate the meaning, for the participating women, of the lived experience of receiving notification about an abnormal Pap smear result. The data are presented in terms of two themes: Pap smear for routine and recurrent confirmation of health and unexpected and ambiguous communication about Pap smear results. The findings are discussed as an unintentional transition from confirmation of health to liminality. Whereas medical diagnosis has been discussed as structuring the inchoate, an abnormal Pap smear did not create order for the interviewed women. On the contrary, the notification of an abnormal Pap smear created disorder as the women had expected to be confirmed as healthy but instead neither health nor disease were confirmed or excluded. Even 'simple' technology is shown to have an ontological dimension, with the ability to transform daily taken-for-grantedness of ourselves as primarily healthy to (potentially) unhealthy. PMID- 15043602 TI - A grief ignored: narratives of pregnancy loss from a male perspective. AB - This paper, utilising a narrative approach, aims to describe the experiences of men whose partner had experienced pregnancy loss, based on data from Northern Ireland. The methodology was based upon observation within pregnancy loss self help groups and in-depth interviews with 14 men who attended the groups. The study also included interviews with 32 midwives and nurses, with the intention of examining attitudes within the medical context towards bereaved fathers. The impact of pregnancy loss on male partners has been largely overlooked in academic research. When a baby dies before birth the loss can be devastating for fathers yet, very often, the world that surrounds them tends to discount their loss, and emotional support and cultural rituals that are normally available to other bereaved individuals are often absent for this group of men. Previous research has shown that men are expected to be emotionally strong in order to support their partner. The present study will show that the perception that men have only a supportive role in pregnancy loss is unjustified, as it ignores the actual life world experiences of the men, and the meanings they attach to their loss, in what may be a very personal emotional tragedy for them where they have limited support available. The study uncovered several recurring themes including self-blame; loss of identity; and the need to appear strong and hide feelings of grief and anger. There is consideration of the need for hospital staff and the wider community to acknowledge the male partner's grief as being a valid response to the bereavement suffered. PMID- 15043603 TI - Medical acupuncture in Germany: patterns of consumerism among physicians and patients. AB - Since the (re) emergence of heterodox medicine across the Western world, there have been numerous interpretations of this phenomenon by the social sciences. Heterodox patients were said to be active consumers holding postmodern values, while heterodox physicians were described as heretics. Medical doctors taking up heterodox medicine were criticised for acting in their own financial interests. To examine these notions, we selected the most prominent heterodox mode of treatment in the German healthcare system: acupuncture. Twenty-six semi structured interviews with medical acupuncturists and their patients were conducted. On the physicians' side, we analyse their styles of practice. To what extent is acupuncture incorporated into biomedical models? Were there any doctors who completely converted to Chinese ideas about health and illness? The patients' activities before and during treatment are addressed. What made them choose acupuncture? How thoroughly did- and do- they collect information on heterodox treatment? The patients' perception of their relationship with their physicians and the decision-making processes during the consultations are also examined. Finally, we argue that while some modes of heterodox medicine resemble parallel forms of general practice, acupuncture tends to become a medical speciality in which physicians tailor their practice to the individual patient's (perceived) demands. From the patients' perspectives, a passive rather than active form of consumerism emerges, involving ideas on medical services that closely correspond to classical modernity. PMID- 15043606 TI - Quantifying daily hemodialysis. AB - Nearly all published reports and clinical studies of hemodialysis solute kinetics are confined to thrice-weekly dialysis schedules. Over the past 40 years, clinical experience with dialysis treatments given three times per week has expanded enormously, but it was not until the Hemodialysis (HEMO) study results were revealed that nephrologists became fully aware of the limits of usefulness of infrequent dialysis. In light of continued reports of improved quality of life and survival with daily dialysis, it appears that the limits of thrice-weekly dialysis may be extended when treatments are given more often. Analysis of solute kinetics during and between dialyses supports the notion that a more frequent schedule delivers more efficient dialysis and that methods can be developed to allow a comparison of risks among patients treated 3-7 days per week. One such method, based on the concept of solute seclusion, suggests that at the currently established minimum standard dose, approximately 50% of the improvement in solute control afforded by seven treatments per week is achieved by increasing the frequency to four treatments per week. The same model shows that seven treatments per week afford an improvement in solute control that is approximately 80% as effective as continuous dialysis. These conclusions are similar to those derived from a completely different model based on peak concentration toxicity. Neither of these models has been clinically tested, so caution must be advised when treating individual patients. PMID- 15043607 TI - The London, Ontario, Daily/Nocturnal Hemodialysis Study. AB - The London Daily/Nocturnal Hemodialysis Study, a prospective, comparative, nonrandomized study, directly compared outcomes of quotidian (daily) hemodialysis patients with conventional thrice-weekly hemodialysis patients. Patients were assigned to either daily (short-hours) hemodialysis (n = 11) or nocturnal hemodialysis (n = 12) and followed up for 5-36 months; all data were directly compared with matched control patients receiving conventional hemodialysis (n = 22). Outcomes evaluated were adequacy (urea kinetics), nutrition, anemia management, blood pressure and volume control, calcium/phosphorus control, and patient quality of life. In addition, a detailed economic analysis was undertaken. The study showed that both quotidian hemodialysis regimens are more effective than conventional hemodialysis in improving weekly urea clearance. Significant clinical improvements were seen with quotidian therapy in the areas of nutrition (short-hours daily), blood pressure (both), volume control (short hours daily), calcium/phosphorus control (nocturnal), and quality of life (both). A nonsignificant trend for improvement in anemia management was suggested. The economic analysis showed substantial savings in annualized cost per quality adjusted life-year in changing from conventional hemodialysis (carried out in center, in satellite units, or at home) to home quotidian hemodialysis. The substantial clinical benefits of home quotidian hemodialysis, combined with the economic advantage shown by this study, clearly justify its expansion. (The details of this study have recently been published in 11 articles in the American Journal of Kidney Diseases[2003;42(suppl 1)]. PMID- 15043608 TI - Quality of life and daily hemodialysis. AB - Preliminary findings indicate that daily hemodialysis positively impacts patients' energy/fatigue and other uremic and intradialytic symptoms. In addition to improvements in perceived symptoms, improvements in patients' perceived physical and psychosocial functioning have been reported. These findings have come from small series of patients, however, and may reflect an increased attention effect. Confirmation of preliminary findings and identification of changes in other quality of life outcomes await an adequately powered randomized clinical trial. Sleep quality, sexual functioning, and cognitive functioning are quality of life dimensions that may be impacted by daily hemodialysis but about which there is limited information in the preliminary data that exist. Understanding relationships among different levels of quality of life outcomes associated with daily hemodialysis requires consideration of emotional and psychological variables such as burden, depression, and satisfaction with care that may intervene between treatment and the quality of life outcomes that patients report. Deriving health utilities relevant to patient experience on different daily hemodialysis therapies, analyzing longitudinal quality of life outcomes reported by patients on daily hemodialysis, and investigating the effectiveness of daily hemodialysis for specific patient subgroups are research agendas that can provide information needed to facilitate treatment decision making in which quality of life has an important role. PMID- 15043609 TI - Cardiovascular effects of frequent intensive hemodialysis. AB - Cardiovascular disease remains the leading cause of morbidity and mortality for patients with end-stage renal disease (ESRD). Frequent intensive hemodialysis (short daily hemodialysis [2 hours per session, six sessions per week] and nocturnal home hemodialysis [6 hours per session, five to six sessions per week]) has recently gained increasing popularity as an alternative to conventional hemodialysis (4 hours per session, three sessions per week). There is an emerging body of evidence that frequent intensive hemodialysis offers superior uremic toxin clearance, blood pressure control, and other cardiovascular outcomes. The goals of the present review are to systematically evaluate the available evidence in blood pressure control and cardiovascular outcomes in ESRD and the achievable changes after converting from conventional dialysis to frequent intensive hemodialysis, and to provide possible physiological explanations to account for these important changes of potent markers of adverse events in this patient population. PMID- 15043610 TI - Short daily hemodialysis and nutritional status in patients with chronic renal failure. AB - Malnutrition is a frequent complication in hemodialysis patients and is associated with increased mortality and morbidity. Interventions such as oral or intravenous nutritional supplements have often failed to improve nutritional status. We report here the effect that daily dialysis, practiced in our center since 1997, has had on nutritional parameters. Seventeen patients treated with conventional hemodialysis (4-5 hours, three times per week, for 9.6 +/- 8.4 years) were converted to short daily hemodialysis (2-2.5 hours, six times per week, for a mean of 39.1 +/- 23.5 months). Dietary, anthropometric, and biochemical evaluations were performed during conventional hemodialysis, after 1 year on short daily hemodialysis (sDHD(year)), and at the end of follow-up (sDHD(end)). Daily protein intake increased from 1.21 +/- 0.27 g/kg/day with conventional hemodialysis to 1.51 +/- 0.47 g/kg/day at sDHD(year) and 1.51 +/- 0.37 g/kg/day at sDHD(end). Energy intake increased from 33.6 +/- 9.5 kcal/kg/day to 38.3 +/- 10.9 kcal/kg/day at sDHD(year) and 39.4 +/- 9.4 kcal/kg/day at sDHD(end). The normalized protein equivalent nitrogen appearance (nPNA) increased from 1.19 +/- 0.34 g/kg/day with conventional hemodialysis to 1.34 +/- 0.43 g/kg/day sDHD(year) and 1.37 +/- 0.37 g/kg/day sDHD(end). Biochemical indicators also increased: serum albumin increased from 40.2 +/- 3.3 g/L to 44.5 +/- 4.6 g/L and 45.1 +/- 4.1 g/L, and prealbumin increased from 0.32 +/- 0.06 g/L to 0.38 +/- 0.09 g/L and 0.36 +/- 0.09 g/L, respectively. These improvements were accompanied by an increase in body weight from 62.0 +/- 10.6 kg on conventional hemodialysis to 64.3 +/- 10.2 kg at sDHD(year) and 65.5 +/- 9.7 kg at sDHD(end). All the changes between conventional hemodialysis and short daily hemodialysis were statistically significant. Increased frequency is more important than increased dialysis dose. Short daily hemodialysis appears to be a suitable method to improve nutritional status in dialysis patients. PMID- 15043611 TI - Sleep apnea and daytime sleepiness in end-stage renal disease. AB - Sleep disorders are common in patients with end-stage renal disease (ESRD). The prevalence of sleep apnea is 10 times greater in patients with ESRD than in the general population. Although sleep apnea is not improved by conventional modes of dialysis, it is corrected by nocturnal hemodialysis, which provides a new and unique model to study its pathophysiology in this patient population. In addition to causing sleep disruption and impairment of daytime function, sleep apnea may also increase the cardiovascular morbidity and mortality that is commonly found in patients with ESRD. "Pathological" daytime sleepiness is found in 50% of patients with ESRD. Although its pathogenesis has been related both to sleep apnea and periodic limb movements, it has also been attributed to a variety of metabolic factors, including the severity of uremia. Further research is required to evaluate the impact of sleep disorders on the clinical outcome of patients with ESRD. PMID- 15043612 TI - Development of a nocturnal home hemodialysis (NHHD) program for children. AB - Whereas the success of nocturnal home hemodialysis (NHHD) as a treatment modality for adults has been clearly demonstrated, there is very little experience with NHHD in children. There are genuine concerns related to patient safety, as well as a potentially overwhelming burden on the parents of children, while supervising NHHD. Each of these issues has been considered in the development of the NHHD program for children in Toronto. The steps involved in the development of this program are outlined. The experience of Simonsen (4), that NHHD is feasible and provides excellent dialysis for children, is confirmed. PMID- 15043613 TI - Reducing hemodialysis costs: conventional and quotidian home hemodialysis in Canada. AB - The cost of hemodialysis is a concern as the prevalence of end-stage renal disease (ESRD) increases. While home hemodialysis has been described as less expensive than in-center hemodialysis, the proportion of patients performing home hemodialysis has been declining. In Canada, there is increasing interest in daily (or quotidian) forms of home hemodialysis, such as nocturnal hemodialysis and short daily hemodialysis. We reviewed the recent Canadian experience with the costs of these modalities and identified four descriptive costing studies: two whole-program comparisons of home conventional hemodialysis and unmatched in center hemodialysis patients, and two comparing quotidian home hemodialysis and matched in-center hemodialysis patients. All costs are listed as per patient-year in 2003 U.S. dollars. In the two whole-program analyses, conventional home hemodialysis was less expensive than in-center hemodialysis (36,840 USD versus 100,198 USD[p < 0.001] and 34,466 USD versus 58,959 USD[p < 0.001]). Reductions in categories such as staffing and overhead are likely due to the modality, while reductions in medications and hospital admissions may be due to differences in the patient mix. The savings for quotidian home hemodialysis are significant, but less striking. In the two matched analyses, nocturnal hemodialysis cost less than in-center hemodialysis (48,656 USD versus 59,476 USD [p = 0.006]), and costs decreased by 8,046 USD in those converted to short daily hemodialysis and by 14,341 USD in those converted to nocturnal hemodialysis, while increasing by 2,521 USD in those who remained on in-center hemodialysis. Home conventional hemodialysis is less expensive than in-center hemodialysis. Much of these savings are lost when dialysis frequency increases; however, quotidian home hemodialysis remains less expensive than in-center hemodialysis. PMID- 15043614 TI - The direction of end-stage renal disease reimbursement in the United States. AB - In the 1960s, care of the end-stage renal disease (ESRD) patient changed. Either through transplantation or intermittent hemodialysis ESRD patients were able to live longer. The Gottschalk Committee issued the "Report of the Committee on Chronic Kidney Disease" to the Bureau of Budget. This report established that hemodialysis and kidney transplantation were the standard of care for the treatment of ESRD patients. Section 2991 of the Social Security Amendment of 1972 provided that patients with ESRD who qualified for Medicare were entitled to receive dialysis and/or transplantation nationwide. The first Medicare hearing held in 1975 regarding the Medicare ESRD program found that the cost of the program was much greater than had been predicted in 1972. Ever since 1975, accelerating cost has been a concern of the government. Quality of care for ESRD patients has been a major focus of the renal community and the Centers for Medicare and Medicaid Services (CMS) over the past 10 years. Consensus for entitlement has remained, but the staggering cost of the program as a whole has become a major problem that must be addressed. Since 1978, Congress and the CMS have passed laws and developed demonstration projects to improve quality of care and reduce costs in the Medicare ESRD program. Kt/V, urea reduction ratio (URR), and hemoglobin levels have all improved, but hospital admissions have remained unchanged and mortality rates have increased. Do the renal community and CMS need to rethink how they will improve the quality of care for ESRD patients and reduce costs in the Medicare ESRD program? PMID- 15043615 TI - An international registry to compare quotidian dialysis regimens with conventional thrice-weekly hemodialysis: why, how, and potential pitfalls. AB - Thrice-weekly hemodialysis is the most commonly used form of renal replacement therapy, yet it is associated with unacceptably high morbidity and mortality. Attempts to improve outcomes for hemodialysis patients by increasing their per session dose of dialysis have recently proven unsatisfactory in the multicentered Hemodialysis (HEMO) study. Interest has thus turned to increasing dialysis frequency. Short daily and long nocturnal dialysis, which are typically performed 6 days per week, are gaining acceptance and are associated with significant improvements in secondary outcomes, including nutrition, left ventricular hypertrophy, hypertension, anemia, and calcium-phosphorus balance. Studies to date have not been adequately powered to detect the survival benefits that these changes may confer. Large-scale randomized studies are planned, but will likely not answer the survival question for several years. Until this issue is resolved, funding policies are unlikely to change, confining current dialysis patients to potentially suboptimal therapy. By capturing data from current and future daily dialysis patients using an international registry, a survival benefit might be demonstrated more quickly. Such a project will soon be undertaken by the London Daily/Nocturnal Study Group with endorsement from the International Society for Hemodialysis and the U.S. National Institutes of Health. This database will also provide useful descriptive data that will help develop methodologies in this growing field. Historically the interpretation of dialysis registry data has been plagued with various methodological problems. These are briefly reviewed, and some potential solutions and necessary precautions are discussed. PMID- 15043616 TI - Evidence-based medicine in the dialysis unit: a few lessons from the USRDS and the NCDS and HEMO trials. AB - This short review argues for the importance of evidence-based medicine in the evolution of improved dialysis care. The National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK) has supported two major randomized clinical trials in dialysis, the National Cooperative Dialysis Study (NCDS), completed in 1981, and the Hemodialysis (HEMO) study, completed in 2002, and supports a major body of ongoing observational research through the U.S. Renal Data System (USRDS). The main results from the two randomized clinical trials and their implications for clinical practice and subsequent research are discussed here. The importance of observational research in the formulation of hypotheses and improvement of quality of care is examined. A brief historical perspective is provided on the critical examination of dialysis care in the late 1980s that occurred following completion of the NCDS. Finally, the rationale for continued investment in dialysis research and the plans of NIDDK to launch studies of more intensive dialysis regimens are discussed. PMID- 15043617 TI - Predicting treatment dose for novel therapies using urea standard Kt/V. AB - Calculation of urea standard Kt/V (stdKt/V) as a dose measure for guiding novel hemodialysis or hemofiltration therapy prescription is complex since this parameter depends on the magnitude of posttreatment urea rebound. We propose here a two-step procedure for calculating urea stdKt/V from single-pool urea Kt/V values (spKt/V) determined from serum urea concentrations in pretreatment and posttreatment blood samples. First, the dependence of urea stdKt/V on equilibrated Kt/V (eKt/V) was derived from a fixed-volume single-pool model. Second, an empirical equation for predicting urea eKt/V from urea spKt/V values was determined using multiple linear regression and available data during 4-hour hemodialysis, 2-hour hemodialysis, and 2-hour hemofiltration treatments. This empirical (rate/dose) equation is likely more robust for novel therapies than other equations derived from only data during conventional (4-hour) hemodialysis treatments. The combination of these formulas allowed construction of nomograms for calculating urea stdKt/V from spKt/V during novel therapies. These principles were further illustrated by calculating the predicted treatment dose for daily (six times per week) hemofiltration therapy required to achieve a given urea stdKt/V. PMID- 15043618 TI - Rationale and experience with short daily hemofiltration. AB - Patients with end-stage renal disease (ESRD) have morbidity and mortality rates that are much higher than those found in the general population. The nonphysiologic nature of thrice-weekly hemodialysis may be partially responsible for the poor outcomes in this patient population. In response to the problems encountered with conventional hemodialysis schedules, a number of investigators have explored potential solutions to the problem. One such solution is to increase the time and/or frequency of hemodialysis. With this approach, ESRD patients have been observed to have better anemia management and blood pressure control, as well as improvements in quality of life. Hemodialysis is based on diffusion of solutes across a semipermeable membrane such that only small molecules are readily removed. Therefore a second solution has been to utilize hemofiltration instead of hemodialysis. Hemofiltration utilizes convective clearance of molecules, resulting in an improvement in the removal of larger solutes, much like the glomerular basement membrane of the kidney. Despite the accumulated evidence that thrice-weekly hemofiltration adds to the benefits seen with increasing hemodialysis time and/or frequency, the use of hemofiltration has been limited by the available technology, cost, and low access blood flows. However, current technological advances in equipment and access creation are addressing these problems. With these advances, a number of investigators have also explored the potential benefits of daily hemofiltration as a treatment option for ESRD. The preliminary results suggest that improvements in blood pressure, beta2-microglobulin (beta2M) levels, and quality of life are possible. PMID- 15043619 TI - The Aksys personal hemodialysis system. AB - The Aksys personal hemodialysis system empowers patients to dialyze where, when, and how they want. It allows them to easily do the best dialysis--daily hemodialysis--in the safest place--at home--by simplifying operation (almost anyone can do it), doing the drudgery work of dialysis (the patient is the master, not the slave of the dialysis machine, giving patients their lives back), needing no bulky disposables (patients have their homes back), reducing emergencies during dialysis, and performing immediate fluid resuscitation on demand. PMID- 15043620 TI - Baxter Aurora dialysis system. AB - With the recent focus on the benefits of more frequent dialysis, the Baxter Aurora dialysis system provides maximum flexibility for therapy prescription, including short daily treatments, long nocturnal treatments, hemodialysis, hemofiltration, and online hemodiafiltration, all in a compact, reliable, easy to use system. A self-prompting touch screen user interface mounted on a movable arm provides for comfortable operation, whether sitting and standing. An automatic treatment setup mode facilitates easy treatment setup. Complex menus are eliminated by the use of a hardware key that automatically selects only the prescribed options during power up, eliminating all menus associated with nonprescribed functions and modalities. This prevents the user from becoming confused or accidentally altering the dialysis treatment. Prior to dialysis the instrument goes through an automatic self-test that confirms the operation of internal systems. The screen will dim when there is no action that the patient needs to attend to on the instrument. After dialysis, press the disinfect button and the instrument disinfects itself and shuts off. For patient safety, the "disinfect" menus are not available during dialysis. The instrument can also be programmed to automatically start and rinse at a set time. For remote treatment monitoring, the instrument connects to the Internet. The Aurora records information about the machine's technical status, providing a record of instrument history for easy servicing. The Aurora is a flexible platform that provides the desired renal therapy with ease of use and proper support for the hemodialysis patient when combined with Baxter's 24-hour infrastructure and support. PMID- 15043621 TI - Bellco Formula Domus Home Care System. AB - There are certain characteristics in a dialysis machine that would be desirable for use in home and limited care environments. These features relate to safety, ease of use, consideration of physical space, and reliability. The Bellco Formula Domus Home Care System was designed to meet all these requirements. Bellco's philosophy of patient treatment centers on global biocompatibility. This is evident in the design of the Formula Domus Home Care System. It has the smallest hydraulic fluid pathway of any dialysis machine on the market. Formula is capable of preparing ultrapure dialysate. The ultrafiltration measurement mechanism, the patented Coriolis flow meter, measures the mass of the dialysate, not the volume. For this reason it is the only dialysis machine that detects actual backfiltration, not just the theoretical possibility of it based on transmembrane pressure. The Coriolis flow meter also ensures that dialysate flow is a true single pass. The operator interface is a single window operating control. It is possible to select up to 14 different languages. There is an online help key to assist patients with troubleshooting. Programmable start-up and shutdown times save time for the patient. Formula is the only dialysis machine to offer a backup battery feature. Formula is capable of communicating with any software available. The focus on global biocompatibility ensures the best quality dialysis treatments for a population of patients who will likely remain on dialysis for a longer period of time than conventional dialysis patients. PMID- 15043622 TI - The Fresenius Medical Care home hemodialysis system. AB - The Fresenius Medical Care home dialysis system consists of a newly designed machine, a central monitoring system, a state-of-the-art reverse osmosis module, ultrapure water, and all the services associated with a successful implementation. The 2008K@home hemodialysis machine has the flexibility to accommodate the changing needs of the home hemodialysis patient and is well suited to deliver short daily or prolonged nocturnal dialysis using a broad range of dialysate flows and concentrates. The intuitive design, large graphic illustrations, and step-by-step tutorial make this equipment very user friendly. Patient safety is assured by the use of hydraulic systems with a long history of reliability, smart alarm algorithms, and advanced electronic monitoring. To further patient comfort with their safety at home, the 2008K@home is enabled to communicate with the newly designed iCare remote monitoring system. The Aquaboss Smart reverse osmosis (RO) system is compact, quiet, highly efficient, and offers an improved hygienic design. The RO module reduces water consumption by monitoring the water flow of the dialysis system and adjusting water production accordingly. The Diasafe Plus filter provides ultrapure water, known for its long term benefits. This comprehensive approach includes planning, installation, technical and clinical support, and customer service. PMID- 15043623 TI - The Gambro system for home daily dialysis. AB - Safety and reliability have been the main emphasis when developing our system for home daily dialysis. The AK 95 is part of a comprehensive system of appropriate products consisting additionally of a silent water treatment module, an ultrafilter, and a range of dry disposables for dialysis fluid preparation and disinfection. The dialyzer can be selected from a family of synthetic, biocompatible filters, both low and high flux. To complete the system, a modern data management tool for online or off-line surveillance and multilingual training manuals in both conventional format as well as animated software are available. PMID- 15043624 TI - The Allient dialysis system. AB - The Allient is a dialysis system that combines various technologies to allow dialysis to be performed at sites outside of dialysis units (intensive care unit [ICU] or home) with ease and safety. A sorbent column regenerates dialysate, removing toxins and providing ultrapure dialysate from only 6 liters of tap water. The use of the sorbent column eliminates the need for costly and complex water purification systems. The Pulsar Blood Movement System provides blood flow at constant negative or positive pressure through single-lumen or dual-lumen accesses, maximizing blood flow rate while eliminating bothersome pressure alarms. Ultrasonic flow monitors control the operation of the pump and ensure adequate blood flow during each dialysis treatment. A completely disposable blood tubing and dialysate circuit eliminates the need for sterilization of the machine. The Allient should make dialysis in the ICU or home setting much more practical, reducing training requirements and increasing safety. PMID- 15043625 TI - The NxStage System One. AB - Given the results of recent randomized controlled trials as well as staffing and budget challenges that today face many institutions across North America, a novel therapeutic approach is likely necessary to enable improvements in clinical outcomes for renal failure patients. The NxStage System One was developed to address these challenges. The system is an innovative, flexible device that delivers hemodialysis, hemofiltration, and/or ultrafiltration therapies to patients with renal failure or fluid overload. The unique characteristics of this system include a highly automated system design with a drop-in cartridge to facilitate training and simple operation; portable size and independence from dedicated infrastructure to minimize practical barriers to where therapy may be administered; use of high-quality premixed treatment fluids to enable capture of the potential clinical benefits of fluid purity without the hassles of local water treatment; and wide operating ranges to allow clinician flexibility in patient therapy prescriptions. In both the chronic and acute care environments, the System One presents clinicians with a new platform for delivering patient therapy improvements within real-world constraints. PMID- 15043626 TI - Modification of the peritoneoscopic technique of peritoneal dialysis catheter insertion: experience of an interventional nephrology program. AB - Bowel perforation is a well-recognized complication of peritoneal dialysis catheter insertion and is associated with increased morbidity and cost of medical care. In this article we describe our 2-year experience (August 2001-October 2003) with a modified peritoneoscopic technique of peritoneal dialysis catheter insertion to minimize the incidence of bowel perforation. Seventy patients underwent 82 consecutive peritoneal dialysis catheter insertions using the innovative technique. The modified technique is very similar to the traditional peritoneoscopic procedure except for the following differences. To gain access to the peritoneal cavity, a Veress insufflation needle (Ethicon Endo-Surgery Inc., Cincinnati, OH) is utilized instead of the trocar. In contrast to the sharp tip of the trocar, the Veress needle has a blunt, self-retracting end. In addition, the Veress needle is only 14 gauge as opposed to the 2.2 mm diameter of the trocar. Upon introduction of the Veress needle into the abdominal cavity, two "pops" are discerned similar to the trocar. After introduction, 400-500 cc of air are infused and the needle is removed. The infusion of air creates a space between the peritoneal surface of the anterior abdominal wall and the bowel loops. At this point, the cannula with trocar is inserted into the space created. The rest of the steps of the procedure are the same as the traditional peritoneoscopic technique. Utilizing the innovative technique, all 82 catheter insertions were performed successfully without a single bowel perforation. No other complications except for catheter migration (n = 2) were noted. The extra cost of the needle (35 USD) should be viewed in the context of the costs associated with management of a bowel perforation. Large-scale studies are needed to confirm the superiority of this innovative technique over the traditional peritoneoscopic insertion found in our case series. In the interim, however, the increased morbidity and cost associated with bowel perforation calls for logical measures to be taken to avoid this dreaded complication. PMID- 15043628 TI - Painful Achilles tendon and its treatment. PMID- 15043629 TI - Physical activity and adipose tissue metabolism. PMID- 15043630 TI - Fat metabolism in exercise--with special reference to training and growth hormone administration. AB - Despite abundance of fat, exclusive dependency on fat oxidation can only sustain a metabolic rate corresponding to 50-60% of VO(2max) in humans. This puzzling finding has been subject to intense research for many years. Lately, it has gained renewed interest as a consequence of increased obesity and physical inactivity imposed by Western lifestyle. Why are humans so poor at metabolizing fat? Can fat metabolism be manipulated by exercise, training, diet and hormones? And why is fat stored in specialized adipose tissue and not just as lipid droplets inside muscle cells? In the present review, human fat metabolism is discussed in relation to how human fat metabolism is designed. Limitations in this design are explored and examples of different designs for fat metabolism from animal physiology are included to illustrate these limitations. Various means of manipulating fat metabolism are discussed with special emphasis on exercise, training, growth hormone (GH) physiology and GH administration. It is concluded that fat stores, non-esterified fatty acids (NEFAs) availability and enzymes for fat oxidation can be increased substantially. However, it is almost impossible to increase fat oxidation during endurance exercise at higher intensities. It seems that, for some reason, the human being is far from optimally designed for fat oxidation during exercise. Acute GH administration has several unexpected effects on fat and carbohydrate metabolism during aerobic exercise, and future research in this area is likely to provide valuable information with respect to GH physiology and the regulation of fat and carbohydrate metabolism during aerobic exercise. PMID- 15043632 TI - Arthroscopic posterior cruciate ligament reconstruction: one- vs. two-tunnel technique. AB - As the techniques of arthroscopic surgery have evolved, so have the possibilities of reconstructing the posterior cruciate ligament (PCL). However, the indications for this procedure have been debated. We describe the results of reconstructing the PCL in 16 consecutive patients with chronic knee instability due to isolated PCL deficiency using either a patella bone-tendon-bone (pBTB) graft with proximal fixation in one femoral tunnel or double bundle hamstring grafts in two femoral tunnels. The patients were followed prospectively and had an additional evaluation after median 35 (25-51.5) months by a blinded observer. The objective stability and subjective scores increased significantly from the preoperative condition. There were no statistically significant differences in outcome between using a pBTB graft in one femoral tunnel compared with semitendinosus/gracilis grafts in two femoral tunnels. We conclude that arthroscopic reconstruction of isolated PCL ruptures improves the activity level of patients and relieves symptoms. In our study, no difference in outcome between the two techniques could be found. PMID- 15043631 TI - Preliminary results of colour Doppler-guided intratendinous glucocorticoid injection for Achilles tendonitis in five patients. AB - BACKGROUND: It is debated as to whether Achilles tendonitis (AT) has an inflammatory component. The intratendinous hyperaemia demonstrated with colour Doppler has been interpreted as neovascularisation. Glucocorticoid injection around the tendon is a common therapeutic procedure. HYPOTHESES: The intratendinous hyperaemia seen with ultrasound (US) colour Doppler represents an inflammatory background. Glucocorticoid injections will be effective if administered inside the tendon where the inflammation seems to be. STUDY DESIGN: An uncontrolled, prospective study with a minimum follow-up of 3 months. METHODS: Six tendons in five patients were evaluated with grey-scale US and colour Doppler before and after US-guided intratendinous glucocorticoid injection. Pain at rest and at activity was evaluated on a visual analogue scale. RESULTS: With colour Doppler all tendons had intratendinous flow. Pain and colour Doppler activity decreased during a mean follow-up of 182 days (range 92-309 days). One tendon relapsed after 199 days. CONCLUSION: Intratendinous glucocorticoid injections seem to have a marked effect on both symptoms and colour Doppler findings, which may be taken as an indication of an inflammatory component in the disease. Colour Doppler adds significant information to grey-scale US with regard to diagnosis, location and follow-up of AT. PMID- 15043633 TI - Fresh meniscal allograft transplantation and autologous ACL/PCL reconstruction in a patient with complex knee trauma following knee dislocation--a case report. AB - Instability of the knee joint, particularly in combination with the loss of one meniscus, regularly leads to the early development of arthritis. This paper describes the case of a 19-year-old male with ruptures of the anterior (ACL) and posterior cruciate ligament (PCL) along with the loss of the medial meniscus due to knee dislocation. Combined, time-delayed reconstruction of both the ACL and PCL and the allogenic fresh meniscal transplantation of the medial meniscus without bone plugs were performed. The control arthroscopy performed 6 months post-transplantation revealed good vitality and integration of the grafts as assessed both macroscopically and histologically. A small portion of the posterior horn had to be refixated, and the anterior horn was atrophic. At 24 months after trauma and 13 months following meniscal transplantation, the patient achieved a Lysholm score of 88 points and clinical examination indicated a stable knee. Fresh meniscal allograft transplantation, in combination with autologous ACL and PCL reconstruction, constitutes--in specialized centers--an alternative treatment option for complex trauma of the knee joint with loss of a meniscus. PMID- 15043634 TI - Gender, muscle, and velocity comparisons of mechanomyographic and electromyographic responses during isokinetic muscle actions. AB - The purpose of this study was to examine the responses of peak torque (PT), mean power output (MP), mechanomyographic (MMG) and electromyographic (EMG) amplitude and mean power frequency (MPF) of the vastus lateralis (VL), rectus femoris (RF), and vastus medialis (VM) in males and females during maximal, concentric isokinetic muscle actions. Subjects performed maximal leg extensions at 60 degrees s(-1), 120 degrees s(-1), 180 degrees s(-1), 240 degrees s(-1), 300 degrees s(-1), 360 degrees s(-1), 420 degrees s(-1), and 480 degrees s(-1). No gender differences were observed, but there were muscle-specific differences for the patterns of MMG MPF, EMG amplitude, and EMG MPF. The MP and MMG amplitude increased to 180-240 degrees s(-1), plateaued, and then decreased to 480 degrees s(-1). MMG MPF for the VL and VM remained unchanged to 300 degrees s(-1), but then increased to 480 degrees s(-1). The EMG amplitude for the RF and EMG MPF for the VL decreased across velocity. Overall, these findings indicated that there were muscle-specific, velocity-related differences in the associations among motor control strategies (EMG amplitude and MPF) and the mechanical aspects of isokinetic muscular activity (MMG amplitude and MPF). PMID- 15043635 TI - A 10-year study of snowboard injuries in Lapland Sweden. AB - Snowboard injuries in a Swedish ski area were evaluated from 1989 to 1999. All injured skiers (alpine, telemark, snowboarders) who sought medical attention at the local Medical Center within 48 h of the accident, were asked to answer an injury form. Physicians assessed and treated the injured skiers. There were a total of 1775 injured skiers; 568 injured snowboarders mean age 19 years. The female/male ratio was 34/66%, the injury rate 3/1000 skier days, three times higher than that of alpine skiers. The skill level of the injured snowboard riders improved during the period. The fall/run ratio of the beginners was higher (1.0) and their risk behavior lower (3.9 on visual analogue scale 1-10) in comparison to the advanced riders (0.4 and 6.6, respectively). Injuries were in 54% located to the upper extremity, 35% were wrist/lower arm injuries. Beginners had significantly higher frequency of lower arm/wrist injuries (46%), than average (32%) and advanced riders (20%). The most frequent single diagnosis was wrist/lower arm fracture (20%). Advanced riders tend to have more head/neck injuries than beginners, 17% vs. 13% (NS). Thus, with elevated skill level the injury pattern changed. For injury prevention, wrist guards and helmets are recommended for snowboard riders. PMID- 15043637 TI - Meeting the educational needs of special populations. AB - We evaluated the Advanced Placement (AP) program from the point of view of intellectually precocious youth and their subsequent educational-vocational outcomes, analyzing normative and idiographic longitudinal data collected across 30 years from 3,937 participants. Most took AP courses in high school, and those who did frequently nominated an AP course as their favorite. Students who took AP courses, compared with their intellectual peers who did not, appeared more satisfied with the intellectual caliber of their high school experience and, ultimately, achieved more. Overall, this special population placed a premium on intellectual challenge in high school and found the lack of such challenge distressing. These findings can inform contemporary educational policy debates regarding the AP program; they also have general implications for designing and evaluating educational interventions for students with special needs. PMID- 15043638 TI - Eye movements do not reflect retrieval processes: limits of the eye-mind hypothesis. AB - This research investigated whether eye movements are informative about retrieval processes. Participants learned facts about persons and locations, and the number of facts (fan) learned about each person and location was manipulated. During a subsequent recognition test, participants made more gazes to high-fan facts than to low-fan facts, and gazes to high-fan facts had a longer duration than gazes to low-fan facts. However, there was no relation between the order in which items were fixated and the relative effect of person or location fan. The effect of person and location fan on gaze duration also did not differ with whether it was the person or location being fixated. A model assuming that the process of retrieval is independent of eye movements was successfully fit to the data on the distribution of gaze durations. According to this model, the effect of fan on number of gazes and gaze duration is an artifact of the longer retrieval times for high-fan facts. PMID- 15043639 TI - Dream rebound: the return of suppressed thoughts in dreams. AB - People spent 5 min before sleep at home writing their stream of thought as they suppressed thoughts of a target person, thought of the person, or wrote freely after mentioning the person. These presleep references generally prompted people to report increased dreaming about the person. However, suppression instructions were particularly likely to have this influence, increasing dreaming about the person as measured both by participants' self-ratings of their dreams and by raters' coding of mentions of the person in written dream reports. This effect was observed regardless of emotional attraction to the person. PMID- 15043640 TI - Don't stand so close to me: the effects fo self-construal in interpersonal closeness. AB - Three studies investigated the effects of self-construal activation on behavior conducive to interpersonal proximity. Study 1 revealed that compared with control participants, participants who were primed with the independent (or personal) self sat further away from where they anticipated another person would sit in a waiting room. Results of Study 2 indicated that participants primed with the interdependent (or social) self sat closer to the anticipated other person than did those primed with the independent self. Finally, Study 3 used the chronic self-construal of participants to predict the seating distance in dyadic settings. Results showed that greater independence of participants' self construals was associated with greater spatial distance during the interaction. Together, the studies provide clear evidence that self-construal activation automatically influences interpersonal behavior as reflected in the actual distance between the self and others. Results are discussed in terms of the functions and motives connected to self-construals. PMID- 15043641 TI - Why the sunny side is up: association between affect and vertical position. AB - Metaphors linking spatial location and affect (e.g., feeling up or down) may have subtle, but pervasive, effects on evaluation. In three studies, participants evaluated words presented on a computer. In Study 1, evaluations of positive words were faster when words were in the up rather than the down position, whereas evaluations of negative words were faster when words were in the down rather than the up position. In Study 2, positive evaluations activated higher areas of visual space, whereas negative evaluations activated lower areas of visual space. Study 3 revealed that, although evaluations activate areas of visual space, spatial positions do not activate evaluations. The studies suggest that affect has a surprisingly physical basis. PMID- 15043642 TI - Vantage point in traumatic memory. AB - Persons with posttraumatic stress disorder (PTSD) spontaneously recalled the traumatic event that led to their condition from either a field or an observer vantage point. In the former case, participants recollected the event as if they were seeing it again through their own eyes; in the latter case, the event was recalled from the perspective of a detached spectator. Analysis of the two types of recollections revealed marked differences in their contents. Whereas field memories afforded richer accounts of the affective reactions, somatic sensations, and psychological states that participants experienced during the focal trauma, observer memories contained more information about participants' physical appearance and actions and the spatial layout of the traumatic scene. Observer trauma memories were also experienced as less emotional and anxiety provoking than field trauma memories. The discussion focuses on the clinical implications of these findings and prospects for future research on traumatic memory. PMID- 15043643 TI - Near-death experiences and the temporal lobe. AB - Many studies in humans suggest that altered temporal lobe functioning, especially functioning in the right temporal lobe, is involved in mystical and religious experiences. We investigated temporal lobe functioning in individuals who reported having transcendental "near-death experiences" during life-threatening events. These individuals were found to have more temporal lobe epileptiform electroencephalographic activity than control subjects and also reported significantly more temporal lobe epileptic symptoms. Contrary to predictions, epileptiform activity was nearly completely lateralized to the left hemisphere. The near-death experience was not associated with dysfunctional stress reactions such as dissociation, posttraumatic stress disorder, and substance abuse, but rather was associated with positive coping styles. Additional analyses revealed that near-death experiencers had altered sleep patterns, specifically, a shorter duration of sleep and delayed REM sleep relative to the control group. These results suggest that altered temporal lobe functioning may be involved in the near-death experience and that individuals who have had such experiences are physiologically distinct from the general population. PMID- 15043644 TI - Amygdala responses to emotionally valenced stimuli in older and younger adults. AB - As they age, adults experience less negative emotion, come to pay less attention to negative than to positive emotional stimuli, and become less likely to remember negative than positive emotional materials. This profile of findings suggests that, with age, the amygdala may show decreased reactivity to negative information while maintaining or increasing its reactivity to positive information. We used event-related functional magnetic resonance imaging to assess whether amygdala activation in response to positive and negative emotional pictures changes with age. Both older and younger adults showed greater activation in the amygdala for emotional than for neutral pictures; however, for older adults, seeing positive pictures led to greater amygdala activation than seeing negative pictures, whereas this was not the case for younger adults. PMID- 15043645 TI - We saw it all along: visual hindsight bias in children and adults. AB - We traced the developmental origins and trajectory of the hindsight bias. Three-, 4-, and 5-year-old children and adults identified gradually clarifying images of degraded common objects on a computer. Half the time, observers did not know in advance what the object would become. The rest of the time, observers knew the object's identity in advance and estimated when a naive same-age peer would identify the clarifying object. In two experiments, children and adults demonstrated hindsight bias by using advance knowledge to overestimate their same age peers' ability to identify the objects. The magnitude of this bias declined across age in one experiment, but remained relatively stable over age in the other experiment. These findings link developmental psychology and adult cognitive science. PMID- 15043646 TI - The basic laws of conditioning differ for elemental cues and cues trained in compound. AB - The cue-duration effect (i.e., longer cues result in less conditioned responding than shorter cues) was examined as a function of whether cues were trained alone or in compound. Compound (AX) or elemental (X) cues of either long or short duration were paired with the unconditioned stimulus. In testing with X alone, the cue-duration effect was observed with elementally trained cues, but not with compound cues. Instead, stronger responding resulted from training with long compound cues relative to short compound cues (i.e., a reversed cue-duration effect). Moreover, an overshadowing effect (i.e., decreased responding due to compound conditioning) was observed when conditioning was conducted with a short cue, but compound conditioning resulted in enhanced responding when it was conducted with a long cue (i.e., reversed overshadowing). These findings are consistent with other recent demonstrations that some laws of learning that apply to elementally trained cues do not similarly apply to cues trained in compound. PMID- 15043647 TI - Inhibition of return from stimulus to response. AB - In a standard inhibition-of-return (IOR) paradigm using a manual key-press response, we examined the effect of IOR both on the amplitude of early sensory event-related brain potential (ERP) components and on the motor-related lateralized readiness potential (LRP). IOR was associated with a delay of premotor processes (target-locked LRP latency) and reduced sensory ERP activity. No effect of IOR was found on motor processes (response-locked LRP latency). Thus, IOR must arise at least in part from changes in perceptual processes, and, at least when measured with manual key presses, IOR does not arise from inhibition of motor processes. These results are consistent with the results of attention-orienting studies and provide support for an inhibition-of-attention explanation for IOR. PMID- 15043648 TI - Does color synesthesia pose a paradox for early-selection theories of attention? AB - P.M. is a synesthete who experiences colors when viewing alphanumeric symbols. Her search for a target differing from distractors by a synesthetic color feature takes the form of a pop-out search. Thus, it would seem that synesthesia can occur preattentively. However, discrepancies between the regression functions of response times observed in target-present trials and target-absent trials, and the fact that fast response times occur only when the target is within a few degrees of visual angle from fixation, indicate that P.M.'s synesthesia does not occur preattentively, but rather is within the focus of attention. We conclude that synesthesia is a genuine perceptual phenomenon that can have substantial influence on visual processing. PMID- 15043649 TI - Keeping it short: a comparison of methods for brief picture presentation. AB - Research has shown that backward masking is a powerful tool for studying unconscious mental processes. Whereas studies have traditionally presented stimuli using cathode-ray tube (CRT) monitors or mechanical shutters together with slide projectors, recent studies (mainly in functional magnetic resonance imaging, fMRI) have begun to use methods based on liquid crystal displays (LCDs) and thin-film transistor (TFT) technology. However, because of differences in technology, all methods may not be equally suited for masking. When methods were compared for their accuracy in presenting pictures at short durations, LCD and TFT presentations had poor accuracy, but shutter and CRT presentations had better accuracy. Because CRTs interfere with the imaging process in fMRI, we recommend the use of mechanical shutters. However, our results may not generally apply to all displays, so we advise researchers to validate the presentation parameters of their displays. The procedure described here may be useful for that purpose. PMID- 15043650 TI - Dread risk, September 11, and fatal traffic accidents. AB - People tend to fear dread risks, that is, low-probability, high-consequence events, such as the terrorist attack on September 11, 2001. If Americans avoided the dread risk of flying after the attack and instead drove some of the unflown miles, one would expect an increase in traffic fatalities. This hypothesis was tested by analyzing data from the U.S. Department of Transportation for the 3 months following September 11. The analysis suggests that the number of Americans who lost their lives on the road by avoiding the risk of flying was higher than the total number of passengers killed on the four fatal flights. I conclude that informing the public about psychological research concerning dread risks could possibly save lives. PMID- 15043651 TI - Color naming: "grue" in the Celtic languages of British Isles. PMID- 15043652 TI - Color naming and sunlight: commentary on Lindsey and Brown (2002). PMID- 15043653 TI - Sunlight and "Blue": the prevalence of poor lexical color discrimination within the "grue" range. PMID- 15043655 TI - Japan's contribution to clinical pediatrics research in the last decade. AB - OBJECTIVE: To determine Japan's contribution to research in clinical pediatrics over the last decade. METHODS: Articles published in highly reputable pediatrics journals from 1991-2000 were accessed through the MEDLINE database. The number of articles which had an affiliation with a Japanese institution were counted for each of the journals and also summed in total. Proportions of randomized controlled trials (RCTs) and case-control/cohort studies among the articles from Japan were also generated and compared with the average for the entirety. In addition, shares of the top-ranking countries were presented along with their trend over time. RESULTS: In total, 20 189 articles were published in the selected seven pediatric journals from 1991-2000. Japan contributed 3.0% of these articles and this contribution was ranked 7th in the world. A negative trend was noticed in Japan's contribution over time but it was not statistically significant (z = -0.40, P = 0.16). RCTs accounted for 7.3% of the total articles, but only 0.34% of those from Japan. CONCLUSIONS: Japan's share of articles in pediatrics research was smaller than that in basic science and some of the other clinical fields. The number of articles from Japan providing a high level of evidence was meager in this field. PMID- 15043656 TI - Determination of total homocysteine in dried blood spots using high performance liquid chromatography for homocystinuria newborn screening. AB - BACKGROUND: The most widely used method for newborn screening for homocystinuria (HCU) is a semi-quantitative bacterial inhibition assay for measuring methionine concentration in dried blood spots (DBS). Because this method has resulted in a number of missed cases due to many factors, we developed a high performance liquid chromatography (HPLC) method with fluorescence detection to measure total homocysteine (tHcy) in DBS which might be useful for newborn screening for HCU. METHODS: One disk of DBS 3 mm in diameter was sonicated in 10 min. The extract was reduced with dithioerythritol and was derivatized with 4-aminosulfonyl 7fluoro-2,1,3-benzoxadiazole before injection into HPLC. RESULTS: This method showed good linearity (r = 0.996), precision (coefficient of variation range 2.7 5%), and excellent correlation coefficient between DBS and serum tHcy, both in control (r = 0.932) and patient samples (r = 0.952). By this method, the mean tHcy concentration in DBS of preterm newborns, full-term newborns, and adults was 1.4 +/- 1.0, 2.5 +/- 1.6, and 4.9 +/- 1.5 micro mol/L, respectively. The mean tHcy DBS concentration in two cases of cystathionine-beta-synthase deficiency and one case of 5,10-methylentetrahydrofolate reductase deficiency was 22.7 +/- 2.88, 29.3 +/- 1.90, and 41.3 micro mol/L, respectively. CONCLUSIONS: The present method, which is rapid, user friendly and reliable, seems applicable to newborn screening of HCU in place of methionine measurement. PMID- 15043657 TI - Nitric oxide inhalation therapy in very low-birthweight infants with hypoplastic lung due to oligohydramnios. AB - BACKGROUND: Although nitric oxide inhalation (iNO) therapy improves arterial oxygenation and reduces the rate of extracorporeal membrane oxygenation in term neonates, the efficacy of this therapy in premature infants is controversial. The objective of the present study was to determine whether iNO therapy improves the survival of very low-birthweight infants with pulmonary hypoplasia due to prolonged rupture of membrane. METHODS: A retrospective comparative study of very low-birthweight infants with pulmonary hypoplasia due to oligohydramnios who had or had not been treated with iNO therapy, was performed (iNO-treated group, eight infants; control group, 10 infants). A neonate was considered to have pulmonary hypoplasia due to oligohydramnios if the following conditions were satisfied: (i) artificial surfactant treatment did not improve the respiratory distress; (ii) prolonged rupture of membrane (PROM) continued for more than 5 days with oligohydramnios; and (iii) sufficient arterial oxygenation did not occur even after giving 100% oxygen, and more than 8 cm H(2)O of mean airway pressure was needed to maintain arterial oxygenation. RESULTS: Nitric oxide inhalation improved arterial oxygenation rapidly and consistently in all eight infants with pulmonary hypoplasia. All eight iNO-treated infants survived longer than 28 days, while five of the 10 control infants died within 24 h of birth (P < 0.05). Before starting iNO, seven of the eight treated infants had shown persistent pulmonary hypertension, which was confirmed by echocardiography. No iNO-treated infant had IVH greater than grade 1, while one control infant had grade 2 IVH. All six long term survivors in the iNO-treated group are developing normally, while only two of the control infants are developing normally as of February 2002. CONCLUSIONS: The majority of the infants with pulmonary hypoplasia due to oligohydramnios had persistent pulmonary hypertension. iNO improved the arterial oxygenation and significantly improved the survival rate. A controlled study to determine whether iNO therapy improves the survival rate of preterm infants with pulmonary hypoplasia due to oligohydramnios is necessary. PMID- 15043658 TI - Sucking behavior at breast during the early newborn period affects later breast feeding rate and duration of breast-feeding. AB - BACKGROUND: One of the objectives of health-workers is to help increase the rate and duration of breast-feeding. It could become a helpful tool if babies sucking behavior at breast in the early neonatal period could predict babies at risk of short-term breast-feeding. The aims of this study are: to determine the prevalence of a range of sucking behaviors at breast in the early neonatal period in Japan; to determine if sucking behavior in the early neonatal period affects the later breast-feeding rate; and to clarify reasons for ceasing breast-feeding for each sucking behavior. METHODS: A total of 1582 mothers of infants aged 6-12 months were surveyed. The questionnaire asked mothers their interpretation from memory of the sucking behavior of their infants in the early neonatal period. Mothers were asked to choose one out of five defined types (barracudas, excited ineffectives, procrastinators, gourmets and resters) and others. The study also included questions about the type of feeding (i.e. full, partial). Finally mothers were questioned about the time of and reasons for ceasing breast-feeding. RESULTS: Sufficient responses to the questionnaires were given by 1474 mothers (93.2%). The remainder of mothers questioned (108, 6.8%) did not provide appropriate answers. The babies whose mothers selected 'Barracudas' as the sucking behavior which best described their babies breast-feeding showed the largest number of full breast-feeding. These babies also displayed the highest breast-feeding rate all the time. The 'Procrastinators' type showed the lowest overall rate of breast-feeding. The Chi-square test revealed that sucking behavior in the early neonatal period affects the breast-feeding rate at 3 and 6 months of age (P < 0.001). The reasons for ceasing breast-feeding observed in all subtypes were perceived shortage of milk volume and elder children's anger and jealousy. CONCLUSION: Based on the sucking behavior at breast in the early neonatal period, it is possible to predict babies at risk for short-term breast feeding. PMID- 15043659 TI - Neonatal urinary tract infections: analysis of the patients and recurrences. AB - BACKGROUND: Early diagnosis and proper treatment, including long-term follow up, are very important for neonatal urinary tract infections (UTI). METHODS: The present study reports the analysis and long-term follow-up results of 71 newborns treated for UTI. RESULTS: Forty-one per cent of patients were preterm babies. Suspected sepsis and hyperbilirubinemia were the main presenting features. Community-acquired and nasocomial UTI accounted for 63% and 37% of cases, respectively. The leading causative agents were Escherichia coli for community acquired UTI and Klebsiella pneumoniae for nasocomial UTI. The urosepsis rate was 5%. Abnormal ultrasonography findings were present in 23% and vesicoureteral reflux was present in 15% of babies. A total of 23% of patients showed renal photopenic areas on dimercaptosuccinic acid scan. The recurrence rate was 28% occurring between 1.5 and 12 months, in particular in the first 6 months. Most of the recurrences developed in patients with no predisposing abnormalities. CONCLUSION: Pediatric nephrologic follow-up of babies experiencing UTI in the neonatal period is very important to identify the predisposing congenital abnormalities and scarred kidneys, to diagnose and to treat the recurrences earlier. PMID- 15043660 TI - Video-fluoroscopic study of swallowing in children with neurodevelopmental disorders. AB - BACKGROUND: Children with neurodevelopmental disorders can have feeding problems. Malnutrition and recurrent aspiration pneumonia can increase the risk of morbidity and mortality. Video-fluoroscopic study of swallowing (VFSS) is essential in understanding the pathological mechanisms involved during swallowing. METHODS: The aim of the present study was to assess the role of VFSS in assessment and management of four children with various neurodevelopmental disorders in a multidisciplinary feeding team. We describe the team approach, with the participation of child neurologist, radiologist with the rehabilitation team including the speech therapist, occupational therapist and dietician, in the assessment and plan of management. RESULTS: Video-fluoroscopic study of swallowing had been useful in assessing the type of swallowing problems with treatment goals targeted to the basic underlying pathophysiological mechanism. CONCLUSION: A child neurologist should be involved in the multidisciplinary oromotor rehabilitation program for neurologically impaired children with feeding problems. PMID- 15043661 TI - Febrile convulsion during the acute phase of Kawasaki disease. AB - BACKGROUND: Although seizures occur in association with meningitis or encephalitis in Kawasaki disease, febrile convulsions in Kawasaki disease are considered to be extremely rare. The aim of the present study is to elucidate the incidence of febrile convulsion in the acute phase of Kawasaki disease, in Niigata City General Hospital, Niigata, Japan. METHODS: The study included 177 patients with Kawasaki disease. Patients ranged in age from 2 months to 10 years (mean age 26.89 +/- 22.44 months). The study included 105 males and 72 females. The clinical records of Kawasaki disease patients were examined retrospectively. RESULTS: Febrile convulsions were not recognized in these 177 patients throughout the course of the disease, despite the presence of a high grade fever and their young age. However, eight of the 177 patients had experienced simple febrile convulsions during other febrile illness except for those with Kawasaki disease. In the acute phase of Kawasaki disease, only two patients showed generalized convulsion associated with prolonged consciousness disturbance and pleocytosis in the cerebrospinal fluid. CONCLUSION: The incidence of febrile convulsions in the acute phase of Kawasaki disease might be extremely low, confirming the results of previous reports. Kawasaki disease is characterized by systemic vasculitis and is sometimes complicated by intracranial vasculitis. The incidence of electroencephalographic abnormalities and pleocytosis in the cerebrospinal fluid is higher in patients with Kawasaki disease. However, the reason why febrile convulsions did not occur in the acute phase of Kawasaki disease remains unknown, despite the presence of central nervous system involvement. PMID- 15043662 TI - Use of laboratory data to identify risk factors of giant coronary aneurysms due to Kawasaki disease. AB - BACKGROUND: Although some laboratory findings are known to be indicators of the risk of giant coronary aneurysm formation among Kawasaki disease patients, an appropriate cut-off point to predict aneurysm formation is not clear. METHODS: One hundred and five patients with giant coronary aneurysms were selected from the 15th and 16th nationwide surveys of Kawasaki disease in Japan. A total of 2936 patients without Kawasaki disease were recruited from a single hospital as a control group. Odds ratios were calculated for six laboratory data with specific values as cut-off points. Receiver operating characteristic (ROC) curves were observed to determine the most appropriate laboratory tests and cut-off points. RESULTS: Hematocrit, leukocyte count, neutrophil proportion, and hemoglobin had one or more peaks of odds ratio for specific cut-off points, but they did not have a clear cut-off point for the predictor according to the receiver operating characteristic curves. Alanine aminotransferase (ALT) increased the risk of giant coronary aneurysms continuously so no clearly appropriate cut-off point was identified. Serum sodium concentration of 135 mEq/L had a peak of odds ratio, and those with <135 mEq/L had the highest odds ratio (4.78). This value seemed appropriate with a sensitivity of 78% and specificity of 57%, although the predictive positive value was as small as 5%. CONCLUSION: The author's propose that a serum sodium concentration of <135 mEq/L at the patient's first visit to hospital may be a predictor of giant coronary aneurysms due to Kawasaki disease. PMID- 15043663 TI - Histological findings after angioplasty using conventional balloon, radiofrequency thermal balloon, and stent for experimental aortic coarctation. AB - BACKGROUND: Use of balloon angioplasty or stent implantation has been reported to be effective in relieving coarctation of the aorta. However, restenosis frequently occurs after balloon angioplasty for native aortic coarctation in small infants, and sometimes develops after stent implantation because of vessel growth. The causes of restenosis remain uncertain. The purpose of this study was to assess the histologic differences in vascular responses to angioplasty using conventional balloon, radiofrequency thermal balloon (RFTB), or stent for experimental aortic coarctation. METHODS: The authors surgically created an aortic coarctation model using 14 puppies. Angioplasty using conventional balloon, RFTB, or stent was performed 1 month after the initial operation. At the acute or chronic phase after angioplasty, the animals were killed and histologic studies were performed. RESULTS: More vascular injuries were noted in the specimens from animals undergoing conventional angioplasty than in those with RFTB or stent. However, neointimal hyperplasia was seen more often after RFTB or stent because of the proliferation of smooth muscle cells from the tunica media, caused by secretion of growth factors. Apoptosis reached a peak 1-2 weeks after angioplasty, regardless of the type of intervention. CONCLUSIONS: The authors conclude that angioplasty with RFTB or stent can provide relatively small injuries in the vessel wall for aortic coarctation, but care must be taken to prevent restenosis caused by intimal hyperplasia, because neointima hyperplasia is more frequent after RFTB or stent. PMID- 15043664 TI - Influence of climate factors on emergency visits for childhood asthma attack. AB - BACKGROUND: Asthma attack shows strong seasonality. The purpose of the present study was to quantify the contribution of climate variables and other seasonal factors on the incidence of emergency visits for childhood asthma in Tokyo, Japan. METHODS: The number of children who visited emergency rooms at Jikei university hospitals in Tokyo during 1998-2002 (5559 visits) was retrieved retrospectively from files from the Department of Pediatrics, and compared with 45 climate parameters from the Meteorological Agency using multiple regression models with a stepwise backward elimination approach. RESULTS: The number of visits (3.7 +/- 3.1) per night increased significantly when climate conditions showed a rapid decrease from higher barometric pressure, from higher air temperature and from higher humidity, as well as lower wind speed. The best-fit model demonstrated that a 22% variation in the number of visits was explained by a linear relationship with 12 climate variables, which increased to 36% after adjusting for calendar month and day of the week. Moreover, when the number of asthma visits was cut off at nine per night, the area under the receiver operator characteristics curve was 0.91 (95% CI: 0.89-0.94) in the multiple logistic regression model using the same variables. CONCLUSIONS: These results suggest that these models might quantify contributions of specific climate conditions and other seasonal factors on the number of emergency visits per night for childhood asthma attack in Tokyo, Japan. PMID- 15043665 TI - Effect of caudal block on stress responses in children. AB - BACKGROUND: The present study was performed during lower abdominal and genitourinary surgery, to assess the effects of caudal block on plasma cortisol, prolactin, insulin and glucose concentrations during and after surgery. METHODS: Thirty male children aged 3-10 years who were scheduled for elective surgery, were selected for the study. The children were premedicated with oral midazolam 0.5 mg/kg. All children received induction with nitrous oxide in oxygen and sevoflurane. The children were randomly allocated into two groups: Group I, control group (n = 15) and group II, caudal group (n = 15). Anesthesia was maintained by face mask with the same agent in both groups. Caudal block was performed with 0.25% bupivacaine 2 mg/kg after induction of anesthesia in the caudal group. Blood samples were obtained after induction of anesthesia (T(0)) in order to measure baseline concentrations of cortisol, prolactin, glucose and insulin. Additional samples were obtained 30 min after the start of surgery (T(1)), and 60 min after the end of surgery (T(2)). RESULTS: All of the basal values (T(0)) were within the normal ranges accepted by Meram Medical Faculty of Selcuk University for children of this age group and there were no differences between the groups (P > 0.05). In both groups, glucose concentration increased at T(1), compared with T(0) and T(2) values (P < 0.05). However glucose concentration was lower in the caudal group than in the control group at T(1) (P < 0.05). In both groups, prolactin concentration increased at T(1), compared with T(0) (P < 0.05). The mean plasma prolactin and cortisol concentration were lower in the caudal group than the control group at T(1) and T(2) (P < 0.05). The mean insulin concentration was lower in the caudal group than the control group at T(2) (P < 0.05). CONCLUSION: These results indicate that caudal anesthesia suppresses the metabolic and endocrine responses to stress associated with lower abdominal and genitourinary surgery in children. PMID- 15043666 TI - Enuresis: prevalence, risk factors and urinary pathology among school children in Istanbul, Turkey. AB - BACKGROUND: Enuresis is a common problem among children and adolescents, and can lead to important social and psychological disturbances. The aim of the present study was to establish the prevalence of enuresis among school children and determine the risk factors associated with this disorder. METHODS: A cross sectional population-based study was conducted in 1576 children. The pupils enrolled in the study were chosen randomly from 14 primary schools located in seven different regions of Istanbul. Data were collected via a questionnaire completed by parents. Enuretic children were invited to the pediatric nephrology outpatient clinic of Cerrahpasa Medical School, Istanbul, Turkey. A detailed history was taken, physical and ultrasonographic examinations, urinalysis and urine culture were performed. The relationship between the prevalence of enuresis and the patients' age, gender, region, the parental educational level and employment status, number of family members, and the family's monthly income were tested by means of chi(2 ) and logistic regression analysis. The comparison between the two enuretic groups (monosymptomatic nocturnal enuresis group vs diurnal enuresis only and diurnal-nocturnal enuresis group) regarding the sociodemographic factors were tested with the chi(2) test and P < 0.05 was accepted as statistically significant. RESULTS: The study group was composed of 1576 school children aged between 6 and 16 years. The overall prevalence of enuresis was 12.4%. When the chi(2) test was used, a significant relationship was found between the prevalence of enuresis and age, educational level of the father, the family's monthly income, and number of family members. However, when logistic regression analysis was applied, there was a statistically significant relationship only between enuresis, and age and number of family members. In the whole group, monosymptomatic enuresis nocturna was found to be more common in boys. When the two enuretic children groups (monosymptomatic nocturnal, diurnal only and nocturnal-diurnal enuretics) were compared with each other regarding gender, parental educational and employment status, and number of family members, statistically significant differences were found. Both maternal and the paternal low educational status were found to be associated with monosymptomatic enuresis nocturna. Likewise, monosymptomatic enuresis nocturna was found to be more common in the children of the unemployed mothers, while diurnal enuresis was more common in the children of unemployed fathers. Nocturnal enuresis was found to be associated with large families. No statistically significant difference was demonstrated between the two groups of enuretics regarding age and family income levels. The rate of urinary abnormalities in the whole group was 7.1%. CONCLUSIONS: Enuresis is a common problem among school children and associated urinary abnormalities are not uncommon. Identification of children at risk is an essential first step before choosing the individualized management for each enuretic child. PMID- 15043667 TI - Child abuse as a result of enuresis. AB - BACKGROUND: Enuresis is a frequent manifestation with important psychological and social consequences. The aim of the present study was to describe the types of child abuse as a result of enuresis, and to investigate the association of child abuse. METHODS: A cross-sectional field study was conducted between March 2001 and August 2001 in the 5-17-year-old age group, to identify risk factors and prevalence of enuresis in the province of Trabzon, Turkey. A face-to-face interview with 889 mothers was carried out. In the questionnaire, the existence, frequency, and risks factors of enuresis were questioned in detail and the parental reactions to the child's enuresis were widely investigated. RESULTS: The prevalence of nocturnal enuresis was 17.9% in the 5-17-year-old age group. It was also found out that of 154 mothers, 86.4% (n = 133) were involved in child abuse of different types. It was found that 40.6% of children who were abused as a result of their enuresis were neglected medically, 42.1% were spanked and 12.8% were beaten. CONCLUSIONS: The high prevalence of enuresis among Turkish children leads to a high frequency of neglect. The high frequency of other types of child abuse in Turkey can be taken as being punishment. PMID- 15043668 TI - Renal resistive indexes and some renal functions in liver cirrhotic children. AB - BACKGROUND: The aim of the present study was to investigate renal vascular resistive changes in children with different stages of liver cirrhosis without obvious renal failure. METHODS: Twenty-nine children (14 girls, 15 boys, mean age 11.6 years) with cirrhosis and 20 healthy children (mean age 10.3 years) were investigated for renal vascular resistance with Doppler ultrasonography, urinary sodium, N-acetyl-beta-D glucosaminidase (NAG) and microalbuminuria excretion. RESULTS: The measurements of renal resistive indexes (RRI) were significantly higher in cirrhotic patients than the control group (0.69 +/- 0.07 vs 0.62 +/- 0.02, P < 0.0001). RRI measurement was found to be increased in decompensated cirrhotic patients than in compensated cirrhotic patients (0.73 +/- 0.05 vs 0.67 +/- 0.08, P < 0.0001). A significant positive relationship was observed between RRI and child score (r = 0.53). Urine NAG/Cr ratio was significantly higher in cirrhotic patients than in the control subjects (P < 0.001). Microalbumin concentrations were increased in the patients with decompensated cirrhosis than in the controls (P = 0.02). Patients with ascites and portal hypertension showed increased RRI values. CONCLUSIONS: We conclude that patients with cirrhosis are at risk of renal deterioration, which can not be detected by serum urea, creatinine, and glomerular filtration rate. The increase of RRI is associated with the progress of hepatocellular disease, and also the development of ascites and portal hypertension. Elevated urinary sodium excretion, elevated urinary NAG/Cr ratio and microalbuminuria might have a prognostic value especially in patients with Child scores> 6. Hence, monitoring RRI is a non-invasive means of studying early renal hemodynamic alteration in childhood cirrhosis. PMID- 15043669 TI - Management for fistula-in-ano with Ginseng and Tang-kuei Ten Combination. AB - BACKGROUND: Non-operative management for patients with fistula-in-ano in infancy using a traditional herbal medicine Ginseng and Tang-kuei Ten Combination (GTTC) is described. METHODS: A total of 22 patients with peri-anal abscess or fistula in-ano were treated with GTTC. Eighteen patients showed no improvement using the conventional non-operative management (frequent curettage of abscess cavity), after which administration of GTTC was commenced. The other four patients received GTTC from the beginning of their treatment. A dose of 0.1-0.2 g/kg of GTTC was given orally twice a day. The period between onset of peri-anal abscess and the commencement of GTTC (pre-GTTC period) was compared to the period until remission after administration of GTTC (post-GTTC period). The relationship between the GTTC dose, age at onset of peri-anal abscess, post-GTTC period, and relapse rates were investigated. RESULTS: Twenty-one patients reached remission, but a 4-year-old boy failed to reach remission and he required fistelectomy. Pre GTTC periods were from 11 days to 3 years (median 50 days) and statistically longer than post-GTTC periods (4-65 days, median 7 days). There was a reverse correlation between the age at onset of peri-anal abscess and post-GTTC period (r = -044, P < 0.05). There was no statistical correlation between the dose of GTTC and post-GTTC period. Five patients showed relapse after discontinuing GTTC, all of them could be resolved with augmentation of the dose. The onset of peri-anal abscess in patients who relapsed were all < 3 months of age. CONCLUSION: Ginseng and Tang-kuei Ten Combination seemed to accelerate the recovery of patients with fistula-in-ano in infancy. PMID- 15043670 TI - Management of thyroglossal duct cysts in children. AB - BACKGROUND: The recurrence rate of thyroglossal duct cysts removed by Sistrunk or other procedures is 4% and 50%, respectively. The aim of the present study was to explain the reasons for recurrence and misdiagnosis. METHODS: Twenty-seven children underwent surgery for thyroglossal cysts and fistulas during 1989-2000. Age, sex, length of history, presentation, preoperative investigations, operative findings, histopathology of the lesion and length of excised hyoid bone, postoperative complications and length of follow-up were recorded. Statistical analysis was performed using Fisher's exact test with a significance level of P < 0.05. RESULTS: Twenty-three patients were primary referrals and four were secondary referrals having had previous surgery with misdiagnosis. The recurrence rate after a Sistrunk procedure was similar to the rate indicated in the literature (3.7%); however, no related special features could be identified such as inflammation (n = 12), perforation at surgery (n = 7), presentation with fistula (n = 4) or previous drainage of abscess (n = 5) (P > 0.05). CONCLUSION: Misdiagnosis is the most common cause of inadequate and inappropriate surgery, leading to recurrence of the lesion. The authors recommend a Sistrunk procedure for all cases of suspected thyroglossal duct cysts. PMID- 15043671 TI - Familial mediterranean fever medicated with an herbal medicine in Japan. PMID- 15043672 TI - Limb body wall defect associated with rare cardiac anomalies. PMID- 15043673 TI - Renal tubular dysgenesis complicated with severe cranium hypoplasia. PMID- 15043674 TI - Congenital multiple myofibromatosis: is it really due to under estrogenic stimulation? PMID- 15043675 TI - Leiomyoma of the diaphragm in a child with Klippel Trenaunay syndrome. PMID- 15043676 TI - Tuberculous brain abscess in a patient with hyper IgE syndrome. PMID- 15043677 TI - Streptococcus pneumoniae-associated arthritis of a lumbar facet joint in a 4-year old boy. PMID- 15043678 TI - Ecthyma gangrenosum combined with multiple perforations of the small intestine associated with Pseudomonas aeruginosa. PMID- 15043679 TI - Unusual cause of vaginal discharge. PMID- 15043680 TI - Cyclosporine A for heavy proteinuria in a child with Henoch-Schonlein purpura nephritis. PMID- 15043681 TI - Endothelial dysfunction in Kawasaki disease: focus on nitric oxide. PMID- 15043682 TI - The expanding horizons of pediatric diabetes. PMID- 15043683 TI - The effect of immunomodulators on prevention of autoimmune diabetes is stage dependent: FTY720 prevents diabetes at three different stages in the diabetes resistant biobreeding rat. AB - BACKGROUND: Autoimmune diabetes of the diabetes-resistant biobreeding (DRBB) rat shares similarities with diabetes in humans and has stages of diabetes that can be controlled and compared. FTY720 is an immunomodulator that has been efficacious in transplant and autoimmune models without inducing an immunosuppressed state. We determined the stages of diabetes that are affected by FTY720 in the DRBB rat. METHODS: Autoimmune diabetes was induced with RT6.1 T cell-depleting antibody and polyIC starting at 4 weeks of age. FTY720 (1 mg/kg/d) was started at day 0, 5, 7, and 14 following the start of depletion. The rats that did not develop diabetes were maintained for 60 d following the last dose of FTY720 before undergoing a second course of depletion. RESULTS: FTY720 starting at day 0, 5, 7, and 14 of depletion prevented diabetes in 100, 100, 50, and 20% of the DRBB rats compared to 0% of the control rats. The surviving rats in the 5 , 7-, and 14-d groups developed diabetes after FTY720 treatment was stopped. Histological examination indicated insulitis in the control rats between day 7 and 11 of depletion and end-stage insulitis by day 18 of depletion compared to negligible insulitis in rats without diabetes. Redepletion in the surviving day 0 rats resulted in development of diabetes in 25% of these rats compared to none of the age-matched controls. SUMMARY: FTY720 can prevent autoimmune diabetes, if administered before and/or during stimulation and expansion of the autoreactive T cells or in the early stages of insulitis. The effectiveness diminishes with each successive stage of diabetes. PMID- 15043684 TI - Continuous glucose monitoring in children with type 1 diabetes: before and after insulin pump therapy. AB - OBJECTIVE: The aim of continuous subcutaneous insulin infusion (CSII) therapy in patients with type 1 diabetes mellitus (T1DM) is to mimic as closely as possible the normal physiologic pattern seen in individuals without diabetes. This study was undertaken to determine the specific areas of improved glycemic control in subjects after initiation of insulin pump therapy and times where further improvement is needed. RESEARCH DESIGN AND METHODS: Eight patients with T1DM (age 7.5-17 yr) wore the Continuous Glucose Monitoring System (CGMS) (Medtronic MiniMed, Northridge, CA, USA) for 3 d before and 3 months after initiation of insulin pump therapy. The CGMS, which measures inter- stitial glucose concentrations every 5 min for a 72-h period, was used to evaluate glucose profiles. Patients entered 4-5 fingerstick blood glucose measurements daily into the sensor for calibration. Detailed logs of food intake, exercise, and hypoglycemic symptoms were also recorded. RESULTS: Hemoglobin A1c (HbA1C) was reduced (p < 0.007) following 3 months of insulin pump therapy. Post-CSII continuous glucose profiles demonstrated an overall improvement in hourly mean glucose over a 24-h period (p < 0.001) as well as a reduction in the area under the curve for glucose (27 +/- 4 prepump vs. 8.6 +/- 1.4 mg/dL/d postpump, p < 0.004). This improvement was a result of an attenuation of the maximal postprandial glycemic excursions. Postbreakfast 349 +/- 24 vs. 267 +/- 16 mg/dL, p < 0.003; lunch 340 +/- 16 vs. 217 +/- 20 mg/dL, p < 0.003. Postdinner average similarly decreased after 3 months of CSII by 22%, p < 0.04. CONCLUSIONS: Pump therapy specifically improved the postprandial glucose excursions in children. PMID- 15043686 TI - Insulin resistance is not due to persistently elevated serum tumor necrosis-alpha levels in small for gestational age, premature, or twin children. AB - BACKGROUND: In pregnancies with small for gestational age (SGA) fetuses, elevated amniotic fluid tumor necrosis factor-alpha (TNF-alpha) levels have been reported. TNF-alpha has been shown to induce insulin resistance in rodents and humans. We hypothesized that an adverse fetal or early neonatal environment for SGA, twin, and premature children leads to persistently elevated TNF-alpha levels that induce insulin resistance in each of these groups. METHODS: The study group consisted of 16 SGA, 14 premature, 53 twin subjects, and the control group of 40 normal subjects (10 short-stature and 30 normal-stature). All subjects were prepubertal and non-obese. Insulin sensitivity (S(I)) was measured in all but the normal-statured control subjects. Fasting plasma TNF-alpha and cortisol levels were measured in all subjects. RESULTS: The study group had reduced S(I)[SGA 18.5 +/- 3, premature 17.8 +/- 2, twin 12.7 +/- 0.7 (x10(-4)/min/ microU/mL)] compared to the short normal control subjects (43 +/- 8 x 10(-4)/min/ microU/mL, p < 0.001). Plasma TNF-alpha levels were lower in the insulin-resistant study group when compared to the control group (2.9 +/- 0.1 vs. 5.0 +/- 0.2 pg/mL, p < 0.001). An association was present between reduced S(I) and low plasma TNF-alpha levels in the study group (p = 0.01, r = 0.4). Fasting plasma cortisol was lower in the study compared to the control group (266 +/- 16 vs. 341 +/- 28 nmol/L, p < 0.01) due to the influence of the twin study subgroup. There was no relationship between plasma cortisol and TNF-alpha levels (p = 0.3). CONCLUSION: SGA, premature, and twin children are insulin resistant and have low plasma TNF-alpha and cortisol levels. We speculate that the mechanism leading to insulin resistance in these subjects is also suppressing plasma TNF-alpha and cortisol concentrations. PMID- 15043685 TI - Redefining the clinical remission period in children with type 1 diabetes. AB - PURPOSE: To redefine the clinical remission period for different aged children receiving the current standard of diabetes care. METHODS: An electronic patient records system was used to identify 552 children newly diagnosed with type 1 diabetes (T1D) from 1997 to 2001 who had an initial hemoglobin A1c (HbA1c) value at the time of diagnosis and at least one other value measured in the ensuing year. The insulin dosage previously used to define the remission period [<0.5 units per kg body weight per day (U/kg/d)] was evaluated for the different aged children. RESULTS: The mean insulin dosages for all age groups were >0.5 U/kg/d by 9 months after diagnosis. The mean HbA1c values were above 8% by 6 months after diagnosis for the 6-9 and the 10-12 yr age groups and by 9 months after diagnosis for the >or=13 yr age group. The percentage of children 30 kg/m(2). It is associated with a number of significant medical conditions, as well as psychological morbidity related to poor body image and the social stigma of obesity. Conservative management is rarely successful in patients with morbid obesity and bariatric surgery is an alternative more likely to produce sustained results. METHODS: To compare the initial results of the Lap Band (Inamed Health, Santa Barbara, CA, USA) procedure when performed by experienced general laparoscopic surgeons, new to the procedure and those achieved by dedicated bariatric practitioners. RESULTS: The results of the present study showed forty-seven per cent excess weight lost at 2 years, 49% excess BMI lost at 2 years. Reoperation rate for band removal, prolapse/slippage was 25.3%. CONCLUSIONS: Acceptable results with Lap-Band are technically achievable by experienced laparoscopic surgeons with a low morbidity and mortality. The results of the present study are inferior to those reported by dedicated bariatric practices who have performed large numbers of this operation. The rate of band slippage was unacceptably high and there was a significant problem with patients being lost to follow up. Possible reasons for this are discussed. PMID- 15043725 TI - Surgical management of obesity using a soft adjustable gastric band. AB - BACKGROUND: Laparoscopically inserted adjustable gastric bands are an effective means of weight reduction for obese patients. METHODS: The Swedish Adjustable Gastric Band was evaluated prospectively in a consecutive series of 273 patients from August 1996 to March 2003. RESULTS: The first 58 patients had their band sited by the 'peri-gastric' technique, with the subsequent 215 using the 'pars flaccida' technique, which provides better posterior fixation of the band. Results from these two groups were considered separately. Prolapse of the stomach through the band was the major complication occurring in 13 of the 58 peri gastric patients (22.4%). Median preoperative body mass index (BMI) was 42.1, falling below 30 by 2 years. In the pars flaccida group there were no instances of prolapse, but the major complication was three cases of band erosion (1.4%). Median preoperative BMI was 42.9 for this group, falling to 32 by 3 years. There was one death in the series due to myocardial infarction. CONCLUSIONS: Results compare favourably with other published adjustable gastric band series. Weight loss with the SAGB appears durable overall with an acceptable complication rate once an appropriate technique is employed. Consistent life-long follow up is necessary to achieve the best result. PMID- 15043726 TI - Obesity is a surgical disease: overview of obesity and bariatric surgery. PMID- 15043727 TI - Surgical risk management: the value of a weekly surgical radio-pathological meeting. AB - BACKGROUND: Clinical risk management in surgery involves organizational practices focused on the prevention of adverse outcomes. The present study evaluates the value of a weekly clinical meeting involving surgeons, radiologists and pathologists as a model of clinical risk management within a general surgical department. METHODS: Data for all meetings conducted over a 6-month period were prospectively collected. Changes to previously reported pathology and radiology results, revisions to patient diagnosis, management and recommendations for subsequent patient care arising from the meetings were recorded. RESULTS: Data were collected for 18 meetings. One hundred and eleven patients were reviewed. Seventy-six patients had previously reported radiological investigations. Sixty five patients had previously reported pathology specimens. Four per cent of radiology reports and 8% of pathology reports were amended at the meeting. Seven per cent of patients received a revised diagnosis. Twenty-seven per cent of patients had suggestions for changes in management. Changes to patient care arose from pathologist input in seven patients, from radiologist input in nine patients and from clinician input in 14 patients. Four patients were recommended for an alternative surgical procedure. Five patients were recommended for cancellation of a planned procedure. Adjuvant chemo/radiotherapy was considered for four patients. Referral to specialist surgeons was suggested for two patients. CONCLUSIONS: Weekly multidisciplinary clinical meetings allow early revision and refinement of patient diagnosis and management. As such, they are an effective pre-emptive incident monitoring system, facilitating regular audit and quality control as part of an overall strategy for surgical clinical risk management. PMID- 15043729 TI - Successful antimicrobial therapy and implant retention for streptococcal infection of prosthetic joints. AB - BACKGROUND: Streptococci cause up to 20% of prosthetic joint infections but this has received little attention in the published literature. METHODS: We reviewed retrospectively our experience with treatment of streptococcal prosthetic joint infections. Patients were followed up for up to 15 years after discontinuation of antimicrobial therapy and up to 8.5 years while on continuous antimicrobial therapy. RESULTS: Eighteen cases were diagnosed between 1984 and 1995. These included one group A, seven group B, one group D, seven group G and one viridans group streptococcal infection as well as one group B and D streptococcal co infection. All were late-onset infections and most (11 of 18, 61%) were acute. Hip and knee joints were equally affected. Six of seven group G streptococcal infections were associated with skin or soft tissue infections. Sixteen patients were treated primarily with antimicrobial agents including 5 days to 6 weeks given intravenously and 2 weeks to 8.5 years given orally. At latest follow up, 10 patients had been off antimicrobial therapy for at least 18 months without relapse, one patient had been off antimicrobial therapy for 7 months without relapse and four infections were successfully controlled with long-term suppressive antimicrobial therapy. One infection was unable to be controlled with antimicrobial therapy. CONCLUSIONS: Our results, and those of others, show that prosthetic joint infections caused by streptococci have a relatively good outcome with primary antimicrobial therapy and, when necessary, drainage, lavage or debridement. Provided the prosthesis is stable and the patient can tolerate long term antimicrobial therapy, this may be an effective alternative to excision arthroplasty. PMID- 15043730 TI - Carotid endarterectomy in octogenarians. AB - BACKGROUND: The role of carotid endarterectomy in octogenarians is unclear as this age group was not included in the major trials of carotid surgery. The aim of the present study was to determine the trends and early outcome of carotid endarterectomy performed in octogenarians at a single institution. METHODS: A retrospective review of all carotid endarterectomies performed between 1990 and 2001 was conducted as part of a clinical audit. Trends in procedure numbers, length of stay, age and early outcome were assessed. The results of octogenarians were analysed separately. RESULTS: Three hundred and fifty-nine carotid endarterectomies were performed in 329 patients including 33 octogenarians. The number of cases and the mean age of patients increased and length of stay decreased during the study period. There was a significant increase in the proportion of octogenarians undergoing carotid endarterectomy (P = 0.03) in the second half of the study period. The combined rate of stroke and death was higher in octogenarians compared with patients under 80 years old, but the difference was not statistically significant (8.8%vs 5.8%; P = 0.59). CONCLUSION: The mean age of patients and the proportion of octogenarians undergoing carotid endarterectomy has increased over the 12-year study period. PMID- 15043731 TI - Comparative study on acute pancreatitis management. AB - BACKGROUND: Guidelines have been published regarding the management of acute pancreatitis by the British Society of Gastroenterology (BSG). The aim of the present paper is to compare the management of patients with acute pancreatitis in a tertiary referral medical centre and a regional health centre in Australia during 2001, evaluate compliance with the published BSG guidelines, and compare our data with those of a similar UK study. METHODS: Patients with a primary diagnosis of acute pancreatitis were identified retrospectively. Eighty-four admissions from the Austin Hospital (AH), a tertiary referral centre, and 83 from The Geelong Hospital (TGH), a regional health centre, were treated in these two hospitals. The histories were collected and examined for compliance with the guidelines recommended by the BSG. We compared our data with the data from the two UK hospitals in a previous study. RESULTS: Only 38% of patients from these two centres had all the investigations performed for severity stratification as recommended by BSG. In other respects, AH and TGH managed these patients with acute pancreatitis according to the recommendations. The overall mortality rate from acute pancreatitis was 3.0%, and within the group of severe acute pancreatitis the mortality rate was 22.7%. 65.5% of patients from AH with gallstone related acute pancreatitis had a cholecystectomy or sphincterotomy and extraction of gallstones within 4 weeks of presentation. There were five re admissions to AH in 2001 due to non-operated gallstone-related acute pancreatitis. In contrast, 84.3% of patients from TGH had definitive treatment within 4 weeks and there were three re-admissions to TGH. CONCLUSION: Overall, both a tertiary referral centre and smaller regional hospital in Australia managed acute pancreatitis according to recently published BSG guidelines. The guidelines emphasized the importance of expertise in hepatopancreatobiliary surgery, availability of intensive care unit/high dependency unit and dynamic CT scanning. The recommendations for definitive treatment of patients with gallstone related pancreatitis within 4 weeks of presentation reduced the morbidity and mortality in this group. Although compliance with the guidelines on investigation for severity stratification of acute pancreatitis was poor, this lack of formal severity assessment did not appear to influence the outcome. PMID- 15043732 TI - Impact of laparoscopic cholecystectomy on hospital utilization. AB - OBJECTIVE: The objective of the present study was to assess the impact of laparoscopic cholecystectomy (LC) and associated endoscopic retrograde pancreatography (ERCP) on hospital utilization. BACKGROUND: Laparoscopic cholecystectomy (LC) has resulted in marked reductions in average length of hospital stay; but population-based studies of hospital utilization have generally not taken into account increased cholecystectomy rates or associated increases in pre and postoperative admissions. METHODS: We conducted a population based study of all residents of Western Australia who underwent cholecystectomy in the period 1980-2000. Record linkage was used to identify pre and postoperative admissions, and to estimate aggregate length of stay per case based on all relevant admissions. We estimated trends in cholecystectomy rates, proportions of cases with related pre and postoperative hospital admissions, average aggregate length of stay per case and total bed utilization per unit of population. RESULTS: The introduction of LC was associated with a sustained increase in rates of cholecystectomy of 25%. Similar increases occurred in the percentage of cases with related preoperative and postoperative admissions. Average length of stay for index admissions declined by nearly 60% compared with 50% for all related admissions. Per capita hospital utilization for index admissions decreased by 45% compared with 38% for index and associated admissions combined, and 32% for all admissions for biliary disease. CONCLUSIONS: Reduced hospital utilization associated with LC was partly offset by increases in pre and postoperative admissions and a sustained increase in cholecystectomy rates. Record linkage is required to assess the true impact of new technologies on hospital utilization. PMID- 15043733 TI - Is palliative resection of the primary tumour, in the presence of advanced rectal cancer, a safe and useful technique for symptom control? AB - INTRODUCTION: At some time, every general surgeon will be faced with the task of trying to decide what to do with a patient who presents with rectal cancer and unresectable distant metastases. How safe is resectional surgery? What sort of palliation may be expected following resection of the primary tumour? In an attempt to answer these questions, the management and outcomes of all patients with rectal cancer and distant metastases, who were primarily referred to the colorectal unit at King Faisal Specialist Hospital were examined. METHODS: All patients who underwent primary surgery for rectal cancer in the presence of metastatic disease were identified. The charts of these patients were examined and their morbidity, mortality and survival were determined. RESULTS: Over an 8 year period 22 patients (average age 54 years) underwent rectal resectional surgery in the presence of metastatic disease. There were 13 men and nine women. The commonest complaint was rectal bleeding. All patients had chest radiographs. Pulmonary metastases were identified in four patients. Nineteen abdominal and pelvic computed tomography scans were performed and eight showed evidence of metastases. Skeletal radiographs in two patients showed evidence of bone metastasis. At operation, intraperitoneal metastases were found in 18 patients. Nine of these were not identified preoperatively. Six patients underwent abdomino perineal resection, nine anterior resection and seven a Hartmann's procedure. Eight patients developed a significant postoperative complication and one died 42 days after surgery. The mean length of hospital stay was 18.6 days. Nine patients received preoperative radiotherapy. Four patients had palliative radiotherapy, two for bony, one for liver and one for peritoneal metastases. Patients were followed up for a mean of 1.1 years. During follow up, 11 returned to the emergency room on 24 occasions. Two patients required readmission. No patient had further rectal bleeding. The mean survival was 1.3 years. CONCLUSION: Patients with rectal cancer and unresectable distant metastases can be successfully palliated by resection of the primary tumour with low morbidity and mortality. The early involvement of a palliative care team facilitates patient management and helps patients enjoy what remains of the rest of their lives at home, in comfort and with good symptom control. PMID- 15043734 TI - Cutaneous melanoma in Caucasian New Zealanders: 1995-1999. AB - BACKGROUND: The aim of the present study was to evaluate the spectrum of cutaneous melanoma in Caucasian New Zealanders. METHOD: Data were obtained from the New Zealand Cancer Registry by way of a computerized search of the melanoma ICD-9 codes from 1995 to 1999. The final database used is for people identifying themselves as European. The denominator population were people stating their ethnicity as European in the 1996 New Zealand Census; all others were excluded. The Cancer Registry Act 1993 made reporting of cancer mandatory. Cancer data before July 1994 are of dubious accuracy. RESULTS: There were 4966 cases of cutaneous melanoma reported in New Zealand between 1995 and 1999 by people identifying themselves as European. The trends and statistically relevant findings will be discussed. CONCLUSION: New Zealand continues to have one of the highest rates of melanoma in the world, with an increasing Breslow thickness of melanoma (P < 0.001) over the 5-year period. Men have a higher rate and deeper melanomas than women (P < 0.001). The incidence of melanoma appears to have reached a plateau over the review period. The far north of New Zealand (Whangarei and further north) had the highest rate of melanoma in New Zealand (59.1/100000, age standardized) and the lowest rate is in Southland (23.5/100000). PMID- 15043735 TI - Modified Limberg flap reconstruction compares favourably with primary repair for pilonidal sinus surgery. AB - BACKGROUND: The present study analyses the results of wide excision with primary closure (PC), wide excision with classical Limberg flap reconstruction (LF) and wide excision with modified Limberg flap reconstruction (MLF) in the surgical treatment of sacrococcygeal pilonidal disease. METHODS: One hundred and sixty-two well-documented patients who were operated on for pilonidal disease and followed for more than 1 year were analysed retrospectively. Group 1 was composed of patients with excision plus PC (n = 78) while group 2 included those with excision plus a LF reconstruction (n = 40), and group 3 included those with excision plus a MLF reconstruction (n = 44). RESULTS: There were no significant differences among the three groups with respect to age, sex distribution, frequency of recurrent disease, or follow-up periods (P > 0.05 for all comparisons). Significant disadvantages regarding postoperative infection rate, mobilization time, discharge from hospital, and time off work were noted for primary closure, compared with both LF and MLF reconstructions. Following a median follow-up period of 4.2 years, 14 recurrences (17.9%) developed in the PC group, three (7.5%) in the LF group, and none (0%) in the MLF group. The zero recurrence rate in the MLF group was significantly lower than that in the PC group (P = 0.003). On the other hand, the recurrence rate in the LF was not found to differ significantly from that in the PC group (P = 0.126). Comparing the LF and MLF groups, none of the surgical end points reached a statistically significant difference (P > 0.05 for all comparisons). CONCLUSIONS: For the surgical treatment of sacrococcygeal pilonidal disease, excision plus a classical or modified Limberg flap reconstruction proved to be superior to excision plus primary closure in terms of infection, mobilization time, discharge from hospital and time off work. Additionally, MLF reconstruction resulted in a statistically lower recurrence rate when compared with PC. PMID- 15043736 TI - Ureteric stenting 25 years on: routine or risky? AB - BACKGROUND: Ureteric stents have been used in urological practice for over 25 years and in many cases have become almost routine. The purpose of the present review is to highlight the uses, complications and risk management issues associated with their use. METHODS: An extensive literature review was conducted and knowledge from past experience was accessed to give a summary of past and current ureteric stent use in urology. RESULTS: Broadly, there are stone and non stone indications for stenting. Complications may range from the commonly experienced 'stent syndrome' to the medico-legal dilemma of the forgotten stent. Risk management must be applied to all uses of stenting to minimize complications and achieve best practice. CONCLUSION: Although almost routine in many areas of urological practice, the complications and implications for risk management of ureteric stenting cannot be ignored. PMID- 15043737 TI - Sexual function after rectal excision. AB - BACKGROUND: Rectal excision is associated with a risk of autonomic nerve damage and associated sexual dysfunction (SD). The evolution of our understanding of the anatomy and physiology of sexual function together with continual refinement of surgery for both benign and malignant disease has led to a decrease in the incidence of SD after rectal surgery. A knowledge of the degree of risk of postoperative SD is important both for the patient and as a benchmark for audit of individual colorectal practice. METHODS: The available literature on the anatomy, physiology and surgical aspects of this topic has been researched through the Medline database. The more recently available data are reviewed in the context of the historical evolution of surgery for benign and malignant rectal disease. RESULTS AND CONCLUSIONS: In the best hands, permanent impotence occurs in less than 2% of patients following restorative proctocolectomy and at a similarly low rate after proctocolectomy and ileostomy. Isolated ejaculatory dysfunction is also numerically a minor problem post operation for benign disease. Patient age is the most important predictor of SD after surgery for rectal cancer. The incidence of permanent impotence remains high (>40%) after abdomino-perineal excision of the rectum (APE) but the continued decline in the use of this operation in favour of low anterior resection (LAR), which carries about half the risk of impotence compared to sphincter ablating surgery, is likely to have resulted in a fall in the absolute number of patients rendered impotent as a result of rectal cancer surgery. Anatomical dissection of the pelvis with preservation of the named autonomic fibres results in a low and predictable rate of sexual morbidity. Surgeons could profitably spend more time with their patients discussing the possible effects of surgery on sexual function. Further research is required to determine the effects of adjuvant therapy for rectal cancer on sexual function. PMID- 15043738 TI - Infarcted intestine: a diagnostic void. AB - The diagnostic approaches to infarction of the small intestine are reviewed in the present paper. The inadequacy of current methods, especially biochemical methods, are highlighted. Additionally, the benefits of a sensitive and specific test of intestinal infarction are discussed. PMID- 15043739 TI - Zymosan induces nitric oxide production by peritoneal mesothelial cells. AB - INTRODUCTION: The production of nitric oxide is an important peritoneal defense mechanism. We have evaluated the effect of various putative stimulants on nitric oxide production by peritoneal mesothelial cells. METHODS: Wistar rats were randomized to either a control group or a peritonitis group (5 mg zymosan intraperitoneally). Groups of five animals were sacrificed at 4, 18, 24, 48 and 96 h after the induction of peritonitis and their peritoneal fluid was harvested for assay. Cultures of peritoneal mesothelial cells were stimulated with lipopolysaccharide, myeloperoxidase, TNFalpha, zymosan, peritoneal fluid from a control animal and peritoneal fluid from a peritonitis animal. Supernatants were collected after incubation for 4, 24 and 48 h for assay. The assay for nitric oxide was based upon the nitrite content of the samples. RESULTS: The intraperitoneal administration of zymosan was associated with an increased production of nitric oxide (NO) when compared with control animals (P < 0.01). In cultures of peritoneal mesothelial cells, zymosan, but not the other putative stimulants, was associated with a marked output of nitric oxide (P < 0.001). CONCLUSION: Zymosan has a direct effect on peritoneal mesothelial cells, which are able to generate nitric oxide in the absence of co-stimulatory molecules. This suggests that it may be possible to use some form of external stimulation to up-regulate the NO response by peritoneal mesothelial cells. PMID- 15043740 TI - Simple, instrument-assisted technique for tying a slip knot: a note of caution. PMID- 15043741 TI - Medical practice and medical education 1500-2001: an overview. AB - Up to the middle of the nineteenth century medicine was practised by a variety of people: physicians, surgeons, apothecaries, bone setters and various irregular practitioners or quacks. Physicians were well educated and learned men who had studied the classics and the works of Galen. The surgeons attended lectures and demonstrations in anatomy. In reality, medical knowledge regarding disease and its management was minimal. The present paper is an overview of the education and training of those who practised in medicine before the subject began to develop into a science. PMID- 15043742 TI - The evolution of the emergency surgeon: the time has come! AB - The challenges now facing the trauma profession are unprecedented. Decreasing reimbursements, malpractice issues, the paucity of operative cases, the difficult work schedule and environment have contributed to fading interest in pursuing a career in trauma. With a steady decline in young surgeons choosing traumatology, a call for redefining the specialty is loudly reverberating throughout the profession. One option that is gaining broad support is to expand the coverage of the trauma surgeon to include all emergency surgical care, thus establishing a new specialist - the 'emergency surgeon'. Although there are no guarantees that widening the spectrum of the specialty will improve the overall financial status of the role, adoption of this new specialty concept will, undoubtedly, enhance the appeal of a dying profession. PMID- 15043743 TI - Emergency thoracotomy and the military surgeon. AB - Emergency thoracotomy might be necessary in battle surgery and might, in an era of superspecialization, be a challenge for the present day military surgeon. He or she might, nonetheless, be the only surgeon in the austere circumstances of a forward field surgical team. This paper provides assistance and guidelines for a military surgeon in such circumstances, including surgical indications and approaches, surgical procedures and techniques. It includes recent advances in this area and principles of 'damage control'. PMID- 15043744 TI - Nasal fractures. PMID- 15043745 TI - Sentinel lymph node biopsy and breast cancer. PMID- 15043747 TI - Minimally invasive surgery for hip replacement. PMID- 15043748 TI - Biliary access loops for intrahepatic stones: results of jejunoduodenal anastomosis. PMID- 15043749 TI - Gallbladder carcinoma presenting as an empyema with Staphylococcus aureus. PMID- 15043750 TI - Neo-adjuvant STI571 therapy for high-risk gastrointestinal stromal tumour. PMID- 15043751 TI - Solid papillary-cystic carcinoma of the pancreas with portal vein tumour thrombus. PMID- 15043754 TI - Prevalence of antimicrobial resistance in bacteria isolated from central nervous system specimens as reported by U.S. hospital laboratories from 2000 to 2002. AB - BACKGROUND: Bacterial infections of the central nervous system, especially acute infections such as bacterial meningitis require immediate, invariably empiric antibiotic therapy. The widespread emergence of resistance among bacterial species is a cause for concern. Current antibacterial susceptibility data among central nervous system (CNS) pathogens is important to define current prevalence of resistance. METHODS: Antimicrobial susceptibility of pathogens isolated from CNS specimens was analyzed using The Surveillance Database (TSN) USA Database which gathers routine antibiotic susceptibility data from >300 US hospital laboratories. A total of 6029 organisms derived from CNS specimen sources during 2000-2002, were isolated and susceptibility tested. RESULTS: Staphylococcus aureus (23.7%) and Streptococcus pneumoniae (11.0%) were the most common gram positive pathogens. Gram-negative species comprised approximately 25% of isolates. The modal patient age was 1 or <1 year for most organisms. Prevalence of MRSA among S. aureus from cerebrospinal fluid (CSF) and brain abscesses were 29.9-32.9%. Penicillin resistance rates were 16.6% for S. pneumoniae, 5.3% for viridans group streptococci, and 0% for S. agalactiae. For CSF isolates, ceftriaxone resistance was S. pneumoniae (3.5%), E. coli (0.6%), Klebsiella pneumoniae (2.8%), Serratia marcescens (5.6%), Enterobacter cloacae (25.0%), Haemophilus influenzae (0%). Listeria monocytogenes and N. meningitidis are not routinely susceptibility tested. CONCLUSIONS: Resistance is commonly detected, albeit still at relatively low levels for key drugs classes such as third generation cephalosporins. This data demonstrates the need to consider predominant resistance phenotypes when choosing empiric therapies to treat CNS infections. PMID- 15043755 TI - Instructions to authors for case reporting are limited: a review of a core journal list. AB - BACKGROUND: Case reports are frequently published in the health care literature, however advice on preparing such reports using the "instructions to authors" pages of journals is alleged to be limited. However, to our knowledge, this has not been formally evaluated. As roles of case reports may vary according to the case and the clinical specialities, one might expect the advice to authors to vary according to journal clinical grouping. METHODS: We surveyed the current advice available to authors of case reports from 'instructions to authors' pages of a core collection of 249 journals ('Hague' list). These were examined and compared for advice or recommendation on writing case reports. Of these, 163 (65%) published case reports and provided instructions on this publication type. Data were extracted on items of style and content of case reports, using a piloted data extraction form. RESULTS: Journals that published case reports were grouped into medical (n = 81, 50%), surgical (n = 38, 23%) and generic or multidisciplinary (n = 44, 27%) categories. There was a difference among the medical, surgical and generic or multidisciplinary journals in the maximum number of words and pages allowed but no difference in the number of figures, tables, references, authors, abstract or synopsis, indexing or key words and consent. Additionally, there was no statistically significant difference among the three different categories of journals regarding the content of the case reports. CONCLUSIONS: Of the journals reviewed, we found that 'instructions to authors' pages provided limited and varied information for preparing a case report. There is a need for consensus, and more consistent guidance for authors of case report. PMID- 15043753 TI - Trophoblast 'pseudo-tumorigenesis': significance and contributory factors. AB - Trophoblast cells of the human placenta proliferate, migrate, and invade the pregnant uterus and its vasculature in order to nourish the developing fetus, in a way that is imitated by malignant tumors. Many similarities exist between embryo implantation and the growth of cancer cells. We begin this article by reviewing decades of studies that have helped unearth the mechanisms that contribute to the tumor-like phenotype of human trophoblast cells. Interestingly, these attributes are only transient in nature, with stringent spatial and temporal confines. The importance of intrinsic molecular controls that effectively circumscribe the extent and duration of trophoblast incursion, becomes increasingly evident in abnormal pregnancies that are characterized by aberrant trophoblast proliferation/invasion. We summarize and discuss the significance of abnormalities in these regulatory mechanisms, and finally, speculate about the use of human trophoblastic cells as model systems for the study of a variety of cellular processes. While on one hand, human placental cells are bestowed with a capacity to proliferate indefinitely and invade extensively, on the other, these cells are also replete with mechanisms to regulate these tumor-like attributes and eventually progress to a senescent apoptotic state. This is therefore, a 'well-behaved' tumor. The comparison in the present review is between the invasive cytotrophoblastic cell type and the tumor cell type. PMID- 15043757 TI - Reliability of sickness certificates in detecting potential sick leave reduction by modifying working conditions: a clinical epidemiology study. AB - BACKGROUND: Medical sickness certificates are generally the main source for information when scrutinizing the need for aimed intervention strategies to avoid or reduce the individual and community side effects of sick leave. This study explored the value of medical sickness certificates related to daily work in Norwegian National Insurance Offices to identify sick-listed persons, where modified working conditions might reduce the ongoing sick leave. METHODS: The potential for reducing the ongoing sick leave by modifying working conditions was individually assessed on routine sickness certificates in 999 consecutive sick leave episodes by four Norwegian National Insurance collaborators, two with and two without formal medical competence. The study took place in Northern Norway in 1997 and 1998. Agreement analysed with differences against mean, kappa, and proportional-agreement analysis within and between groups of assessors was used in the judgement. Agreements between the assessors and the self-assessment of sick-listed subjects were additionally analysed in 159 sick-leave episodes. RESULTS: Both sick-listed subjects and National Insurance collaborators anticipated a potential reduction in sick leave in 20-30% of cases, and in another 20% the potential was assessed as possible. The chance corrected agreements, however, were poor (k < 0.20) within and between groups of National Insurance collaborators. The agreement between National Insurance collaborators and the sick-listed subjects was no better than chance. Neither extended medical information nor formal medical competence increased agreement in cases where modified working conditions might have reduced sick leave. CONCLUSION: Information in medical sickness certificates proved ineffective in detecting cases where modified working conditions may reduce sick leave, and focusing on medical certificates may prevent identification of needed interventions. Strategies on how to communicate directly with sick-listed subjects would enable social authorities to exploit more of the sick leave reduction potential by modifying the working conditions than strategies on improving medical information. PMID- 15043756 TI - Prevalence of hyperuricemia and relation of serum uric acid with cardiovascular risk factors in a developing country. AB - BACKGROUND: The prevalence of hyperuricemia has rarely been investigated in developing countries. The purpose of the present study was to investigate the prevalence of hyperuricemia and the association between uric acid levels and the various cardiovascular risk factors in a developing country with high average blood pressures (the Seychelles, Indian Ocean, population mainly of African origin). METHODS: This cross-sectional health examination survey was based on a population random sample from the Seychelles. It included 1011 subjects aged 25 to 64 years. Blood pressure (BP), body mass index (BMI), waist circumference, waist-to-hip ratio, total and HDL cholesterol, serum triglycerides and serum uric acid were measured. Data were analyzed using scatterplot smoothing techniques and gender-specific linear regression models. RESULTS: The prevalence of a serum uric acid level >420 micromol/L in men was 35.2% and the prevalence of a serum uric acid level >360 micromol/L was 8.7% in women. Serum uric acid was strongly related to serum triglycerides in men as well as in women (r = 0.73 in men and r = 0.59 in women, p < 0.001). Uric acid levels were also significantly associated but to a lesser degree with age, BMI, blood pressure, alcohol and the use of antihypertensive therapy. In a regression model, triglycerides, age, BMI, antihypertensive therapy and alcohol consumption accounted for about 50% (R2) of the serum uric acid variations in men as well as in women. CONCLUSIONS: This study shows that the prevalence of hyperuricemia can be high in a developing country such as the Seychelles. Besides alcohol consumption and the use of antihypertensive therapy, mainly diuretics, serum uric acid is markedly associated with parameters of the metabolic syndrome, in particular serum triglycerides. Considering the growing incidence of obesity and metabolic syndrome worldwide and the potential link between hyperuricemia and cardiovascular complications, more emphasis should be put on the evolving prevalence of hyperuricemia in developing countries. PMID- 15043758 TI - Core I gene is overexpressed in Hurthle and non-Hurthle cell microfollicular adenomas and follicular carcinomas of the thyroid. AB - BACKGROUND: Most of the steps involved in the initiation and progression of Hurthle (oncocytic, oxyphilic) cell carcinomas of the thyroid remain unknown. METHODS: Using differential display and semiquantitative RT-PCR we found, among other alterations, overexpression of the gene encoding the Core I subunit of the complex III of the mitochondrial respiratory chain in a follicular carcinoma composed of Hurthle cells. RESULTS: Similar high levels of Core I gene expression were detected in nine follicular carcinomas (seven with Hurthle cell features), in seven microfollicular adenomas (one with Hurthle cell features) and in one micro/macrofollicular adenoma, in contrast to a lower/normal expression in nine papillary carcinomas (three with Hurthle cell features) and five macrofollicular adenomas (one of which displaying Hurthle cell features). No significative correlation was found between Core I overexpression and the proliferative activity of the lesions. CONCLUSIONS: We conclude that Core I overexpression in thyroid tumours is not associated with malignancy, Hurthle cells or proliferative activity. The pathogenetic mechanism linking Core I overexpression to the microfollicular pattern of growth of thyroid tumours remains to be clarified. PMID- 15043760 TI - The role of the clinical research coordinator--data manager--in oncology clinical trials. AB - BACKGROUND: The purpose of this study was to determine the standard tasks performed by clinical research coordinators (CRCs) in oncology clinical trials. METHODS: Forty-one CRCs were anonymously surveyed, using a four-page self administered questionnaire focused on demographics, qualifications, and professional experience. The survey questions on responsibilities consisted of an ad-hoc 32-item questionnaire where respondents had to rate the frequency of involvement in the listed activities using a 3-point scale. We defined as "standard" a task that was rated as "in all or nearly all trials" by at least half of the respondents. RESULTS: A response rate of 90% (37 out of 41) was achieved after two mailings. Less than half of the respondents had received additional training in oncology, clinical research or Good Clinical Practices (GCP). Overall, all standard tasks performed by CRCs were in the category of "monitoring activities" (those usually performed by a Clinical Research Associate "CRA") and included patient registration/randomization, recruitment follow-up, case report form completion, collaboration with the CRA, serious adverse events reporting, handling of investigator files, and preparing the site for and/or attending audits. CONCLUSIONS: CRCs play a key role in the implementation of oncology clinical trials, which goes far beyond mere data collection and/or administrative support, and directly contributes to the gathering of good quality data. PMID- 15043759 TI - A randomized controlled trial of mycophenolate mofetil in patients with IgA nephropathy [ISRCTN62574616]. AB - BACKGROUND: IgAN is the most common type of glomerulonephritis in the world. Between 15 and 40 percent of adults and children diagnosed with IgAN eventually progress to ESRD. Despite the need for effective treatment strategies, very few RCTs for IgAN have been performed. The most effective therapies for IgAN appear to be corticosteroids, ACEi, and FOS that contain a high concentration of omega 3 fatty acids. While ACEi and FOS are generally well tolerated with minimal side effects, the use of high dose steroids over a long course of therapy is often associated with significant morbidity. OBJECTIVE OF THE STUDY: The objective of the study is to test the hypothesis that treatment with the immunosuppressive agent, MMF, will lead to significant and sustained improvement in urinary protein excretion in patients with IgAN who have been pre-treated (and continue to be treated) with ACEi and FOS compared to a placebo control group of patients receiving comparable doses of ACEi and FOS without MMF. DESIGN: After a three month treatment period with the ACEi, lisinopril and the FOS, Omacor, 100 (2 x 50) patients with IgAN and a urinary P/C ratio > or = 0.6 (males) and > or = 0.8 (females) and an estGFR > or = 40 ml/min/1.73 m2 will be randomized to treatment with either MMF or placebo for one year. All patients will be followed off study drug for a second year, but will continue treatment with lisinopril and Omacor for the two year duration of the study. The primary outcome measure of change in urine P/C ratio will be assessed at the end of years one and two. PMID- 15043761 TI - Selection of informative clusters from hierarchical cluster tree with gene classes. AB - BACKGROUND: A common clustering method in the analysis of gene expression data has been hierarchical clustering. Usually the analysis involves selection of clusters by cutting the tree at a suitable level and/or analysis of a sorted gene list that is obtained with the tree. Cutting of the hierarchical tree requires the selection of a suitable level and it results in the loss of information on the other level. Sorted gene lists depend on the sorting method of the joined clusters. Author proposes that the clusters should be selected using the gene classifications. RESULTS: This article presents a simple method for searching for clusters with the strongest enrichment of gene classes from a cluster tree. The clusters found are presented in the estimated order of importance. The method is demonstrated with a yeast gene expression data set and with two database classifications. The obtained clusters demonstrated a very strong enrichment of functional classes. The obtained clusters are also able to present similar gene groups to those that were observed from the data set in the original analysis and also many gene groups that were not reported in the original analysis. Visualization of the results on top of a cluster tree shows that the method finds informative clusters from several levels of the cluster tree and indicates that the clusters found could not have been obtained by simply cutting the cluster tree. Results were also used in the comparison of cluster trees from different clustering methods. CONCLUSION: The presented method should facilitate the exploratory analysis of big data sets when the associated categorical data is available. PMID- 15043763 TI - Daily rhythm of nociception in rats. AB - BACKGROUND: Many behavioral and physiological variables exhibit daily rhythmicity. Few investigations of the daily rhythmicity in nociception have been conducted, and conflicting results have been obtained. The present study evaluated the daily rhythmicity in nociception in Wistar rats. METHODS: Nociception was investigated by Eddy's hot plate method, tail immersion method, and tail clip method. The latency between the noxious stimulus and the animal's response was recorded as reaction time. Separate groups of rats were tested in 4 hour intervals for 24 hours. RESULTS: There was clear daily variation in response latency. Reaction time was shortest a few hours before lights-on and longest at the light-dark transition. CONCLUSION: Nociception exhibits robust daily rhythmicity in rats. Sensitivity to pain is highest late in the dark phase of the light-dark cycle and lowest at the light-dark transition. PMID- 15043762 TI - Molecular cloning, gene structure and expression profile of two mouse peroxisomal 3-ketoacyl-CoA thiolase genes. AB - BACKGROUND: In rats, two peroxisomal 3-ketoacyl-CoA thiolase genes (A and B) have been cloned, whereas only one thiolase gene is found in humans. The aim of this study was thus to clone the different mouse thiolase genes in order to study both their tissue expression and their associated enzymatic activity. RESULTS: In this study, we cloned and characterized two mouse peroxisomal 3-ketoacyl-CoA thiolase genes (termed thiolase A and B). Both thiolase A and B genes contain 12 exons and 11 introns. Using RNA extracted from mouse liver, we cloned the two corresponding cDNAs. Thiolase A and B cDNAs possess an open reading frame of 1272 nucleotides encoding a protein of 424 amino acids. In the coding sequence, the two thiolase genes exhibited approximately equal to 97% nucleotide sequence identity and approximately equal to 96% identity at the amino acid level. The tissue-specific expression of the two peroxisomal 3-ketoacyl-CoA thiolase genes was studied in mice. Thiolase A mRNA was mainly expressed in liver and intestine, while thiolase B mRNA essentially exhibited hepatic expression and weaker levels in kidney, intestine and white adipose tissue. Thiolase A and B expressions in the other tissues such as brain or muscle were very low though these tissues were chiefly involved in peroxisomal disorders. At the enzymatic level, thiolase activity was detected in liver, kidney, intestine and white adipose tissue but no significant difference was observed between these four tissues. Moreover, thiolase A and B genes were differently induced in liver of mice treated with fenofibrate. CONCLUSION: Two mouse thiolase genes and cDNAs were cloned. Their corresponding transcripts are mostly expressed in the liver of mice and are differently induced by fenofibrate. PMID- 15043764 TI - Alternative splicing affects the function and tissue-specific expression of the human constitutive androstane receptor. AB - BACKGROUND: The constitutive androstane receptor (CAR) plays a key role in the control of drug metabolism and transport by mediating the phenobarbital-type induction of many phase I and II drug metabolizing enzymes and drug transporters. RESULTS: We identified transcripts generated by four different alternative splicing events in the human CAR gene. Two of the corresponding ligand binding domain isoforms demonstrated novel functional properties: First, CAR(SV3), which is encoded by a transcript containing an lengthened exon 7, differentially transactivated target gene promoters. Second, CAR(SV2), which results from the use of an alternative 3' splice site lengthening exon 8, showed ligand-dependent instead of constitutive interaction with coactivators. Furthermore, alternatively spliced transcripts demonstrated a tissue-specific expression pattern. In most tissues, only transcripts generated by alternative splicing within exon 9 were expressed. The encoded variant demonstrated a loss-of-function phenotype. Correct splicing of exon 8 to exon 9 is restricted to only a few tissues, among them liver and small intestine for which CAR function has been demonstrated, and is associated with the induction of CAR expression during differentiation of intestinal cells. CONCLUSION: Due to their specific activities, CAR variant proteins SV2 and SV3 may modulate the activity of reference CAR(SV1). Furthermore, we propose that transcriptional activation and regulation of splicing of exon 9 may be coupled to ensure appropriate tissue- and differentiation state-specific expression of transcripts encoding functional CAR protein. Altogether, alternative splicing seems to be of utmost importance for the regulation of CAR expression and function. PMID- 15043765 TI - Evidence-based pharmacotherapy of schizophrenia. AB - The introduction of the new-generation antipsychotics has changed the way we treat patients with schizophrenia. This article reviews these agents, focusing mainly on the published randomized controlled trials and meta-analyses in which the new-generation antipsychotics are compared with placebo, conventional antipsychotics or with one another. Agents included are risperidone, olanzapine, quetiapine, ziprasidone, sertindole, amisulpride and aripiprazole. Acute-phase and maintenance studies are reviewed, as well as randomized trials for pre psychotic, first-episode schizophrenia and refractory schizophrenia. Finally, specific areas of current clinical interest are dealt with. These are: conventional vs. new-generation antipsychotics, head-to-head comparisons of new generation antipsychotics, and side-effect profiles. PMID- 15043766 TI - The supraorbital keyhole approach with eyebrow incisions for treating lesions in the anterior fossa and sellar region. AB - BACKGROUND: Keyhole surgery has developed since the 1990s as a less invasive therapeutic strategy for intracranial lesions, initially for the treatment of intracranial aneurysms. The purpose of this study was to describe and evaluate the results of surgical treatment of lesions in the anterior fossa and sellar region via a supraorbital keyhole approach using eyebrow incisions. METHODS: Between April 1994 and July 2003, 54 patients with lesions in the anterior fossa and sellar region were operated on via the supraorbital keyhole approach. The surgical results were studied retrospectively and compared with that of patients with lesions at the same locations but treated via a conventional subfrontal approach. RESULTS: No significant difference in curative effect was found between the conventional subfrontal approach and the supraorbital keyhole approach. However, the supraorbital approach required a much smaller skin incision, causing less surgical trauma, while achieving excellent surgical exposure and good recovery. CONCLUSION: The supraorbital keyhole approach using an eyebrow incision is safe, effective, and both suitable and convenient for treating lesions in the anterior fossa and sellar region, with almost no adverse consequences on the facial features of patients. PMID- 15043767 TI - Loss of bone mass after Colles' fracture: a follow-up study. AB - BACKGROUND: Colles' fracture usually associated with osteoporosis is regarded as the predictor of subsequent osteoporotic fracture. However, it is not clear how the local changes of bone mass take place during the course of treatment and whether the changes are related to clinical practice. The objective of the current study was to investigate the local changes of bone mass in patients with Colles' fracture and their possible clinical relevance in a follow-up study. METHODS: The radiograms of the second metacarpal in 64 patients with Colles' fracture were assessed for bone density immediately after fracture, 6 weeks, 6 months and 1 year after fracture, respectively. Functional results were evaluated at one year. RESULTS: Bone mass six weeks after Colles' fracture was significantly decreased without returning to normal at one year though increased bone mass had been identified 6 months after fracture (P < 0.05), (P < 0.01). At one year significant (P < 0.05) or highly significant (P < 0.01) correlations were observed between bone mass indices of metacarpal and functional results, indicating that poor function is associated with lower bone density. Significant differences (P < 0.05) between fracture patterns also suggested that patients with more severe fractures have a more pronounced bone loss. CONCLUSIONS: Bone loss during the course of treatment will have a direct effect upon the prognosis, so different treatment should be proposed for different patterns of fractures. Active exercise should be made to improve the recovery of bone mass. PMID- 15043768 TI - Domestic abuse on women in China before, during, and after pregnancy. AB - BACKGROUND: Some researchers found that partner-perpetrated physical violence increased in frequency and severity during the postpartum period compared with the antenatal period, however, limited data exists describing abuse of women in China. The purpose of this study was to examine patterns of abuse in China before, during, and after pregnancy, and explore possible factors related to abuse. METHODS: A community-based face-to-face survey of a representative group of women who had a child aged 6 to 18 months in 32 communities of Tianjin, Liaoning, Henan, and Shaanxi provinces was carried out between November 1, 2001 and February 28, 2002. RESULTS: The prevalence of domestic abuse (emotional, sexual, or physical) occurring in any period (before, during, or after pregnancy) was 12.6%. The prevalence of abuse during the approximate 9 months of pregnancy (4.3%) was relatively lower compared with the prevalence of abuse during the 12 months before pregnancy (9.1%) and after delivery (8.3%) during the mean 11-month postpartum period studied. Abuse before pregnancy was a strong risk factor for abuse during pregnancy and abuse after pregnancy, and abuse during any previous period was a strong risk factor for subsequent abuse. Many women who suffered abuse of any kind generally experienced multiple acts over time and most acts were not severe. The results of logistic regression analysis showed that the factors associated with abuse during pregnancy included women previously witnessing domestic violence, a poor relationship with the partner, socioeconomic level, alcohol consumption, and smoking. CONCLUSIONS: It is necessary to do in depth training and to raise awareness of partner abuse among all health professionals. Routine screening of abuse in maternity clinics is advocated to decrease the adverse impact of abuse on women and fetuses. PMID- 15043769 TI - Real-time three-dimensional myocardial contrast echocardiography in assessment of myocardial perfusion defects. AB - BACKGROUND: Both real-time three-dimensional echocardiography (RT3DE) and myocardial contrast echocardiography (MCE) are novel imaging techniques. The purpose of this study was to confirm the feasibility and accuracy of RT3DE combined with MCE for quantitative evaluation of myocardial perfusion defects. METHODS: Thirteen dogs underwent ligation of the left anterior descending artery (LAD, n = 6) or distal branch of the left circumflex artery (LCX, n = 7) under general anaesthesia. Three to four ml of a perfluoropropane (C3F8) microbubble contrast agent was injected intravenously to assess the resulting myocardial perfusion defects with a commercially available Philips SONOS-7500 ultrasound system. After removal of the dog hearts, Evans blue dye was injected via the left and right coronary arteries to stain the myocardium at risk. In vitro anatomic measurements of myocardial mass after removal of the animals' hearts were used as controls. RESULTS: Left ventricular (LV) mass determined by RT3DE ranged 36.7 - 68.9 g [mean, (54.6 +/- 9.6) g] before coronary artery ligation, and correlated highly (r = 0.99) with in vitro measurement of LV mass [range, 38.9 - 71.1 g; mean, (55.6 +/- 9.3) g]. There was no significant difference between RT3DE and in vitro measurements of LV mass [range, 36.7 - 68.9 g; mean, (51.3 +/- 12.5) g. Or range, 38.9 - 71.1 g; mean, (53.7 +/- 12.3) g, respectively] and under-perfused mass [range, 0 - 21.4 g; mean, (12.0 +/- 6.9) g. Or range, 0 - 19.8 g; mean, (10.8 +/- 6.3) g, respectively] after the LAD ligation (P > 0.05). Likewise, no significant difference was present between RT3DE and in vitro measurements of LV mass [range, 50.1 - 65.4 g; mean, (57.5 +/- 5.9) g. Or range, 51.5 - 65.8 g; mean, (57.3 +/- 6.4) g, respectively] and under-perfused mass [range, 0 - 25.6 g; mean, (13.3 +/- 9.6) g. Or range, 0 - 22.7 g; mean, (12.8 +/- 8.1) g, respectively] after the LCX ligation (P > 0.05). For all the animals with coronary ligation, LV mass measured by RT3DE ranged 35.9 - 68.6 g [mean, (54.8 +/ 10.0) g] and there was no significant difference between RT3DE and in vitro measurements of LV mass and under-perfused mass (P > 0.05, r = 0.99). Further, the under-perfused mass derived from RT3DE [range, 0 - 25.6 g; mean, (12.7 +/- 8.2) g] correlated strongly with the in vitro measurements [range, 0 - 22.7 g; mean, (11.9 +/- 7.2) g] (r = 0.96). CONCLUSION: RT3DE with MCE is a rapid and accurate method for estimating LV myocardial mass and quantifying perfusion defects. PMID- 15043770 TI - Comparative study of on-pump and off-pump coronary bypass surgery in patients with triple-vessel coronary artery disease. AB - BACKGROUND: Studies on selected patients undergoing off-pump versus on-pump coronary artery bypass surgery have produced inconsistent results, especially in patients with multiple coronary artery disease. This study compared the clinical results of on-pump and off-pump coronary bypass surgery in patients with triple vessel disease. METHODS: A total of 300 consecutive isolated, multiple coronary artery bypass grafting (CABG) patients were assigned to the off-pump coronary artery bypass (OPCAB, n = 150) or CABG with cardiopulmonary bypass (CCABG, n = 150) groups. There were no significant differences regarding degree of angina, history of myocardial infarction or diabetes, and presence of left main coronary artery disease between the two groups. Ejection fraction in the OPCAB group before surgery was lower than in the CCABG group (P < 0.01). In addition, more patients had a history of stroke and abnormal renal function preoperatively in the OPCAB group (P < 0.01). In OPCAB patients, single deep pericardial stay suture with a sling snared down was used to expose the target vessels, along with a stabilizer and a coronary shunt. A Medi-Stim Butterfly Flowmeter was used to measure blood flow through grafts in both groups. RESULTS: No OPCAB patient was converted to the CCABG group. The average numbers of distal anastomoses and the indexes of completeness of revascularization (ICR) were similar in both groups. Postoperative respiratory support time and the volumes of chest tube drainage and of blood transfusions were less in the OPCAB group than in the CCABG group (both P < 0.01). The postoperative incidences of pulmonary dysfunction and renal insufficiency were lower in the OPCAB group than in the CCABG group (both P < 0.05). There were no significant differences between the two groups in mortality and other causes of morbidity (perioperative myocardial infarction, stroke, atrial fibrillation). CONCLUSIONS: OPCAB can be applied to patients with triple vessel coronary artery disease and can achieve similar completeness of revascularization and similar early surgical results, with shorter respiratory support, reduced transfusion requirement, and fewer cases of pulmonary dysfunction and abnormal renal function. PMID- 15043772 TI - Astrovirus infection in young children with diarrhea hospitalized at Beijing Children's Hospital. AB - BACKGROUND: This study was to assess certain epidemiological and clinical characteristics of astrovirus infection in children with diarrhea below 5 years old hospitalized at the Beijing Children's Hospital. METHODS: From September 1999 to August 2001, a total of 288 children with diarrhea below 5 years old hospitalized at Beijing Children's Hospital were studied. Fecal specimens collected from each patient were tested for astrovirus by using enzyme immunoassay (EIA) and further serotyped by reverse-transcription polymerase chain reaction (RT-PCR) using serotype-specific primers. RESULTS: Astrovirus antigen was positive in 9.0% of the patients, and the rate of coinfection with rotavirus was 19.2% among the astrovirus positive patients. The patients with astrovirus infection were at the age of 7 days to 17 months, most (92.3%) of them were less than 12 months. Astrovirus infection occurred mainly between October and March of the following year. Main symptoms included watery stool, vomiting, fever, and dehydration. Increased activity of myocardial enzyme was seen in 4 patients. Of the 22 astrovirus positive specimens, 14 were successfully serotyped by RT-PCR and they were all serotype 1. CONCLUSIONS: The overall incidence of astrovirus infection in young children with diarrhea hospitalized at the Beijing Children's Hospital was 9.0%. The predominant serotype was type 1. Astrovirus may play an important role in viral diarrhea in young children in Beijing, China. PMID- 15043771 TI - Virological and immunological outcomes in HIV-1-infected Chinese patients treated with a combination of Efavirenz and Indinavir for 48 weeks. AB - BACKGROUND: The incidence of HIV-1-related infection diseases and the mortality of AIDS have dramatically decreased since highly active antiretroviral therapy began to be used clinically in China in 1999. And we initiated a second clinical trial using a combination of Efavirenz and Indinavir to observe the effects of the immunoreaction. METHODS: Twenty patients with laboratory-confirmed chronic HIV-1 infection were recruited. Blood samples were collected initially and during the weeks after initiation of treatment. Within 48 hours of blood sampling, peripheral blood plasma and mononuclear cells were separated using routine methods. HIV-1 viral load was measured in thawed plasma samples. Within 48 hours of peripheral blood sampling, CD4(+) and CD8(+) T cell subsets were enumerated. RESULTS: The drug regimen was efficient in reducing HIV-1 plasma viral load and increasing total CD4(+) T cell counts. The percentage of CD4(+) and CD8(+) T cell subsets expressing CD38 and HLA-DR activation markers was positively correlated with plasma viral load and tended to normalize. CONCLUSIONS: The combination of Efavirenz and Indinavir was generally well tolerated and efficient at reducing HIV-1 RNA. Furthermore, the treatment improved the immunological function. PMID- 15043773 TI - Changes of hypothalamus-pituitary hormones in patients after total removal of craniopharyngiomas. AB - BACKGROUND: This paper aimed to elucidate the changes of hypothalamus-pituitary hormones in patients after total removal of craniopharyngiomas. METHODS: A total of 40 patients with craniopharyngiomas received surgery. The levels of triiodothyronine (T3), thyroxine (T4), thyrotropic hormone (TSH), antidiuretic hormone (ADH), and adrenocorticotropin (ACTH) were measured in the 40 patients before surgery and one week after surgery respectively. RESULTS: Twenty-eight patients (70%) had hypothyroidism before surgery, but 38 (95%) had hypothyroidism after surgery (P < 0.01). Twenty-three patients (57.5%) had diabetes insipidus (DI) before surgery and 38 (95%) had DI after surgery (P < 0.001). The pre- and post-operative levels of ADH were (2.49 +/- 0.30) pg/ml and (2.80 +/- 0.29) pg/ml respectively (P > 0.05), whereas those of ACTH were (23.97 +/- 2.69) pg/ml and (15.60 +/- 1.91) pg/ml respectively (P < 0.05). CONCLUSIONS: Hormone deficits after total removal of craniopharyngioma appear to be the common complication of surgery. Hypothyroidism and diabetes insipidus are more frequent after surgery than before surgery. Thyroxine and glucocorticoids should be administered routinely after total removal of craniopharyngioma. PMID- 15043774 TI - Ultrasonographic evaluation of fetal facial anatomy (I): ultrasonographic features of normal fetal face in vitro study. AB - BACKGROUND: Because of lacking skills in scanning the normal fetal facial structures and their corresponding ultrasonic features, misdiagnoses frequently occur. Therefore, we studied the appearance features and improved displaying skills of fetal facial anatomy in order to provide basis for prenatal diagnosis. METHODS: Twenty fetuses with normal facial anatomy from induced labor because of other malformations except facial anomalies were immersed in a water bath and then scanned ultrasonographically on coronal, sagittal and transverse planes to define the ultrasonic image features of normal anatomy. The coronal and sagittal planes obtained from the submandibular triangle were used for displaying the soft and hard palate in particular. RESULTS: Facial anatomic structures of the fetus can be clearly displayed through the three routine orthogonal planes. However, the soft and hard palate can be displayed on the planes obtained from the submandibular triangle only. CONCLUSIONS: The superficial soft tissues and deep bony structures of the fetal face can be recognized and evaluated by routine ultrasonographic images, which is a reliable prenatal diagnostic technique to evaluate the fetal facial anatomy. The soft and hard palate can be well demonstrated by the submandibular triangle approach. PMID- 15043775 TI - Stage-specific expression genes of the Spirometra erinaceieuropaei plerocercoid screened by mRNA differential display technique. AB - BACKGROUND: The stage-specific expression of genes is one of the most characteristics of parasites. It has been found that a lot of genes of Spriometra erinaceieuropaei are specifically expressed in pleroceroid in large amount, but not expressed when the plerocercoid development into adult worm. The study is to screen other stage-specific ecpression genes of plerocercoid of Spirtmetra erinceieuropaei. METHODS: RNA was separately extracted by acid guanidinium thiocyanate-phenol-chloroform from plerocercoids and adult worms of Spirometra erinaceieuropaei, DNA contaminated in the RNA was digested by RNase-free DNase. After the RNA was reverse transcripted to cDNA using T12MA, T12MC, T12MG and T12MT anchor-primers, PCR was done using the same T12MN and one random primer with alpha 35S-dATP in the system. The PCR products were fractionated on an 8% denatured polyacrylamide gel. Differential bands of the plerocercoid found in the gel were cut out, amplified by PCR and sequenced. Northern hybridization was used to identify the stage-specific expression genes. RESULTS: Eleven differential bands were selected from the gel and classified into 3 kinds of gene fragments by hybridization, after they were amplified by PCR. Fragments 1 (238 bp) and 2 (383 bp), were confirmed by Northern hybridization, as being expressed in the plerocercoid. However, fragment 3 (433 bp), was expressed in both the plerocercoid and the adult worm. Data from the 3 gene fragments underwent homological analysis in GenBank. The sequence which was homologous with fragments 1 and 2 was not found, but fragment 3 had high homology with many kinds of 28S rRNA. CONCLUSIONS: The gene expressions of plerocercoids are different from adult worms because they live in different hosts. Two types of different gene fragments from the plerocercoid were found by mRNA differential display technique. PMID- 15043776 TI - Identification of differentially expressed genes in anagen dermal papilla by suppression subtractive hybridization. AB - BACKGROUND: We constructed a cDNA subtractive library of dermal papilla cells (DPCs) in anagen with suppression subtractive hybridization (SSH) technique and clone differentially expressed genes related to DPCs in anagen. METHODS: Total mRNA was isolated from DPCs of anagen and telogen follicles. Moreover, single strand (ss) and double-strand (ds) cDNAs were synthesized in turn using SMART PCR cDNA synthesis technology. ds cDNAs then were digested with Rsa I and divided into two groups, and ligated to the specific adaptor 1 and adaptor 2R, respectively. After cDNAs were hybridized with each other twice and underwent two rounds of nested PCR. PCR products were ligated with arms of T/A plasmid vectors to set up the subtractive library. Selected clones were demonstrated by reverse Northern blot and sequenced. The acquired sequence data were aligned against the Genbank nucleotide database. RESULTS: cDNA subtractive library of DPCs in anagen follicles was set up successfully with high subtractive efficiency. Thirty-five genes were identified in this study with 22 known functional genes and 13 unknown functional genes. CONCLUSIONS: All results confirm the effectiveness and sensitivity of SSH in detecting differentially expressed genes from a small amount of clinical samples. Information about such alterations in gene expression could be useful for elucidating the genetic events in hair follicle growth regulation. PMID- 15043777 TI - Ultrastructural and immunohistochemical studies on Trichomonas vaginalis adhering to and phagocytizing genitourinary epithelial cells. AB - BACKGROUND: Trichomonas vaginalis (T. vaginalis) belongs to a common sexually transmitted disease pathogen causing genitourinary trichomoniasis in both sexes. We investigated the pathogenetic mechanism of genitourinary trichomoniasis. METHODS: Cultured T. vaginalis bodies were injected into the vaginas of rats, or incubated with genitourinary epithelial cells of female subjects, male subjects, and sperm. The ultrastructural and microscopic changes were observed via transmission and scanning electron microscopy and through microscopic histochemistry. RESULTS: Groups of T. vaginalis adhered to PAS positive columnar cells at the surface of stratified epithelium in the middle and upper portions of the vaginas. They also traversed under these cells. The parasites were shown to be PAS, cathepsin D, and actin positive, and they could release hydrolase into the cytoplasm of adhered epithelial cells. In the amebiform T. vaginalis, microfilaments were arranged into reticular formation. Similar phenomena were found during the interaction of T. vaginalis with host cells, both in vitro and in vivo. Usually several protozoa adhered to an epithelial cell and formed polymorphic pseudopodia or surface invaginations to surround and phagocytize the microvilli or other parts of the epithelial cytoplasm. Adhesion and phagocytosis of sperm by the protozoa occurred at 15 - 30 minutes of incubation. Digestion of sperm was found at 45 - 75 minutes and was complete at 90 - 105 minutes. CONCLUSIONS: T. vaginalis tends to parasitize at the fornix of the vagina, because this is the site where columnar cells are rich in mucinogen granules and their microvilli are helpful for adhesion and nibbling. T. vaginalis possesses some invading and attacking abilities. Shape change, canalization, encystation, phagocytosis, digestion, the cell coat, cytoskeleton, and lysosome all play important roles in the process of adhesion. They have two methods of phagocytosis: nibbling and ingestion. Genitourinary epithelium may be injured directly by the digestive action of hydrolases, phagocytosis, and the mechanical action of pseudopodia. PMID- 15043778 TI - The association between paired basic amino acid cleaving enzyme 4 gene haplotype and diastolic blood pressure. AB - BACKGROUND: In a previously identified locus linked to hypertension on chromosome 15q, we identified three blood pressure candidate genes: insulin-like growth factor 1 receptor gene (IGF1R), myocyte specific enhancer factor 2A gene (MEF2A), and paired basic amino acid cleaving enzyme 4 gene (PACE4). In this study, we tested their associations with hypertension using haplotype analysis. METHODS: A total of 288 unrelated individuals, including 163 high diastolic blood pressure (DBP) subjects and 125 normal DBP subjects were enrolled in this case-control study. Twenty single nucleotide polymorphisms (SNPs) in the three genes were genotyped using polymerase chain reaction followed by restriction enzyme digestion. Haplotype analysis was accomplished in the following stages: (1) pair wise linkage disequilibrium test among SNPs on the same gene was performed to explore blocks in which recombination is very unlikely to happen; (2) Estimation Maximization algorithm was applied to estimate haplotype frequencies in each block; (3) the chi-square test was used to examine the specific haplotype difference, and a permutation test was used to examine the overall haplotype profile difference between cases and controls in each block. RESULTS: An estimated haplotype "CCCCG" frequency in the haplotype block on the PACE4 gene was significantly higher in high DBP cases than in controls (P < 0.01). The overall estimated haplotype profile in this block was also significantly different between the cases and the controls (P < 0.001). This association indicates. CONCLUSIONS: This study for the first time demonstrated that PACE4 gene may play an important role in the regulation of DBP. This association indicates that variations influencing DBP resides in or near this genomic region. PMID- 15043779 TI - Evaluation of murine models of permanent focal cerebral ischemia. AB - BACKGROUND: To date murine models of permanent focal cerebral ischemia have not been well characterized. The purposes of this paper were to compare three different permanent middle cerebral artery occlusion (MCAo) models with or without craniectomy, and to identify an ideal mouse model of permanent focal cerebral ischemia. METHODS: Experiments were performed on 45 healthy adult male Kunming mice, weighing 28 to 42 g. The animals were randomly assigned to three groups (n = 15 in every group) based on surgical procedure: MCAo via the external carotid artery (ECA), MCAo via the common carotid artery (CCA), and direct ligation of the middle cerebral artery (MCA). Each day post-ischemia, the animals were scored using an eight-grade neurological function scale, and mortality was also recorded. Seven days post-ischemia, the brains were removed for lesion size determination using triphenyltetrazolium chloride staining. Correlation analysis of lesion volume and neurological score was carried out. RESULTS: Mortality in the group receiving direct MCA ligation was lowest among the three groups, and there was a significant difference between the direct MCA ligation group and the two intraluminal occlusion groups (P < 0.05). In all groups, neurological scores gradually increased with prolongation of ischemic duration, peaking after two days, then gradually decreasing. In the direct MCA ligation group, however, neurological scores were relatively stable. There was a significant correlation between infarct volume and neurological score 7 days after MCAo in every group (all r > 0.7, P < 0.05), suggesting good reproducibility of lesion volume in the three groups, but the infarct volume was more constant in the direct MCA ligation group. CONCLUSION: The direct ligation model of MCAo provides an optimal means of studying permanent focal cerebral ischemia, and is preferable to the models using intraluminal sutures. PMID- 15043780 TI - ROS-mediated ERK activation in delayed protection from anoxic preconditioning in neonatal rat cardiomyocytes. AB - BACKGROUND: The activation of extracellular signal-regulated kinase1/2 (ERK1/2) has been shown to be important signaling pathway in the ischemic preconditioning (IPC) response. Recently, some studies suggest a key role for the mitochondrial ATP-sensitive potassium channel (mKATP) as both a trigger and an end effector of acute and delayed protection of IPC. Hence, this study was undertaken to elucidate the relationship between mKATP and ERK1/2 in the delayed protection mechanism of anoxic preconditioning (APC). METHODS: An APC model was established using cultured neonatal rat cardiomyocytes. Pharmacological agents [diazoxide, 5 hydroxydecanoate (5-HD), 2-mercaptopropionylglycine (MPG), and PD98059] were used to modulate mKATP and ERK1/2 activation. Cellular injury was evaluated by measuring cellular superoxide dismutase (SOD) activity, cell viability, and lactate dehydrogenase (LDH) release. The generation of cellular reactive oxygen species (ROS) and the activation of ERK1/2 were determined at different time points starting from the beginning of preconditioning with anoxia or diazoxide (an mKATP opener). RESULTS: Cell viability and SOD activity in the APC [(81.9 +/- 11.4)%, (13.6 +/- 3.7) U/L] and diazoxide [(79.2 +/- 12.4)%, (16.5 +/- 4.6) U/L] groups were significantly higher than in the anoxia/reoxygenation (A/R) [(42.2 +/ 7.3)%, (8.8 +/- 2.8) U/L] group (all P < 0.01). LDH activity in the APC group [(101.9 +/- 18.9) U/L] and diazoxide group [(97.5 +/- 17.7) U/L] was significantly lower than in the A/R group [(250.5 +/- 43.6) U/L] (all P < 0.01). Both APC and diazoxide simultaneously facilitated intracellular ROS generation and rapid ERK1/2 activation. But the effects of APC and diazoxide were remarkedly attenuated by 5-HP (an mKATP blocker) and by MPG (a free radical scavenger). In addition, the ERK1/2 inhibitor PD98059 also abolished the cellular protective effects induced by diazoxide. CONCLUSION: mKATP may mediate ERK1/2 activation during anoxia preconditioning by generating ROS, which then triggers the delayed protection of APC in rat cardiomyocytes. PMID- 15043781 TI - Changes in myocardial collagen content before and after left ventricular assist device application in dilated cardiomyopathy. AB - BACKGROUND: The purposes of this study were to confirm the changes in myocardial collagen level after left ventricular assist device (LVAD) support in dilated cardiomyopathy (DCM), find the relation between these changes and prognosis, and test a practical method to assess the level of myocardial collagen. METHODS: Left ventricular samples were collected from DCM patients with different prognosis (transplanted group n = 8, weaning group n = 10) at the time when the LVADs were implanted and again during cardiac transplantation (n = 8). The level of neutral salt soluble collagen (NSC) and acid soluble collagen (ASC) was measured by Sircol collagen assay, and that of total collagen and insoluble collagen (ISC) by quantification of hydroxyproline (Hyp). Serum samples were collected from a portion of these patients (transplanted group, n = 6; weaning group n = 7) at the time the LVADs were implanted, 1 month after implantation and on explantation. Circulating concentration of carboxy-terminal propeptide of type I procollagen (P I CP), amino-terminal propeptide of type I procollagen (P I NP), amino-terminal propeptide of type III procollagen (P III NP) and type I collagen telopeptide (I CTP) were measured by the equilibrium type radioimmunoassay. RESULTS: Before LVAD implantation the level of NSC and ISC in the weaning group was higher but ASC in the transplanted group was lower than in the controls (P < 0.05). After LVAD support, the level of total collagen was higher, but ASC was also lower in the transplanted group than in the controls (P < 0.05). In comparison of the pre- and post-LVAD subgroups of the transplanted and weaning groups, all collagen fraction levels before LVAD implantation were lower in the transplanted group than in the weaning group (P < 0.05); but this difference disappeared after LVAD support. Comparison of the pre- and post-LVAD subgroups of the transplanted group showed increased level of NSC and total collagen after LVAD support. The changes of serum peptide concentration showed that P III NP increased constantly in the transplanted group, but P I CP and P I NP increased in the weaning group after LVAD implantation. CONCLUSIONS: The changes in myocardial collagen level as a sign of myocardial interstitial remodeling in DCM are not involved with total collagen but involved with collagen fractions, and they are related to prognosis. The changes of myocardial collagen content and serum procollagen peptide after LVAD support can be regarded as an expression of the reverse of maladaptive myocardial interstitial remodeling. PMID- 15043782 TI - Role of 1,25-dihydroxyvitamin D3 in preventing acute rejection of allograft following rat orthotopic liver transplantation. AB - BACKGROUND: We investigated the role of 1,25-dihydroxyvitamin D3(1,25-(OH)2D3) in preventing allograft from acute rejection following orthotopic liver transplantation. METHODS: A rat orthotopic liver transplantation model was used in this study. SD-Wistar rats served as a high responder strain combination. Recipients were subjected to administration of 1,25-(OH)2D3 at dosages ranging from 0.25 microg.kg(-1).d(-1) to 2.5 microg.kg(-1).d(-1). Survival after transplantation as well as pathological rejection grades and IFN-gamma mRNA, IL 10 mRNA transcription intragraft on day 7, and day 30 post-transplantation were observed. RESULTS: After recipients were treated with 1,25(OH)2D3 at dosages of 0.5 microg.kg(-1).d(-1) or 1.0 microg.kg(-1).d(-1), survivals of recipients were prolonged. Ninety-five percent confidence intervals of survival were 46 - 87 days and 69 - 102 days (both P = 0.0005 vs control group), respectively. On day seven post-transplantation, relative levels of IFN-gamma mRNA transcription were 0.59 +/- 0.12 and 0.49 +/- 0.16, which was higher than the control group (P = 0.005, P = 0.003, respectively). Relative levels of IL-10 mRNA transcription were 0.83 +/- 0.09 and 0.76 +/- 0.09, which was lower than the control group (P = 0.002, P = 0.003, respectively). At a dosage of 0.5 microg.kg(-1).d(-1), the median of pathological rejection grade on day seven and on day thirty post-transplantation were 1.5 and 2.0 in comparison with the CsA-treated group (P = 0.178, P = 0.171, respectively). At a dosage of 0.5 microg.kg(-1).d(-1), the median of pathological rejection grade on day seven and day thirty post-transplantation were 1.5 and 1.5 in comparison with CsA-treated group (P = 0.350, P = 0.693, respectively). CONCLUSION: After each recipient was treated with 1,25-(OH)2D3 at a dosage of (0.5 - 1.0) microg.kg(-1).d(-1), transcription of cytokine intragraft was accommodated effectively and deviated to Th2 type, resulting in alleviation of acute rejection. 1,25-(OH)2D3 can prolong survival of recipient after orthotopic liver transplantation. PMID- 15043783 TI - Changes in the T-cell receptor V beta gene repertoire after allogeneic hematopoietic stem cell transplantation. AB - BACKGROUND: We distinguished graft-versus-host disease (GVHD) from graft-versus leukemia (GVL) effects and to investigate the distribution of T-cell receptor (TCR) V beta gene repertoire in individuals with leukemia before and after allogeneic hematopoietic stem cell transplantation (allo-HSCT). METHODS: Peripheral blood mononuclear cells (PBMC) were obtained from 10 normal individuals, 8 donors and 11 patients with leukemia before and after transplantation. Polymerase chain reaction (PCR) amplification of complementarity determining region 3 (CDR3) of 24 TCR V beta genes was used to examine serial samples of PBMC. The PCR products were further analyzed by genescan to evaluate clonality of T cells. RESULTS: The 24 TCR V beta gene repertoire displayed highly diverse and polyclonal spectratypes in all normal individuals and 4 of 8 donors. Another 4 donors expressed part of the 24 TCR V beta subfamily and 1 donor had oligoclonality. The expressions of the 24 TCR V beta subfamilies were skewed and restricted in 11 leukemia patients before and after transplantation. Some absences of 24 TCR V beta subfamily expression were quite similar between the recipients pro-transplantation and related donors. The number of subfamilies expressed increased over time post-transplantation, but the restricted expressions of the subfamily could last 6 - 30 months after transplantation. All patients with GVHD and some without GVHD exhibited T cell clonal expansion. The expansive T cell clone was distributed in V beta 2-3, 16-17, 18-19, 21 and V beta 23 in patients with GVHD and in V beta 7, 9, 16 and 19 in patients without GVHD. One patient with syngeneic-HSCT (syn-HSCT) had V beta 15 and 16 T cell expansion after transplantation. One patient displayed V beta 18 T cell expansion after donor lymphocyte infusion (DLI). CONCLUSIONS: Normal individuals express the entire 24 TCR V beta gene repertoire and have polyclonal distribution. However, the TCR V beta gene repertoire is only partially expressed in some donors. The TCR V beta gene repertoire is restrictedly expressed in a skew fashion in patients with leukemia before and after transplantation. The number of TCR V beta gene subfamilies increases over time post-transplantation. GVHD and GVL effects may induce the proliferation of T cell clones. Clinical GVL response may be distinguished from GVHD alloreactivity through the host MHC antigen. PMID- 15043784 TI - Study on blocking the leukemia immune escape after BMT by Fas-Fas ligand pathway. AB - BACKGROUND: To investigate if bone marrow transplantation (BMT) with bone marrow mononuclear cells (BMMCs) transducted with murine soluble Fas gene (sFas) using adenovirus vector could block the immune escape of leukemia cells eliminate the residual leukemia cells and reduce their relapse. METHODS: The recombinant adenovirus vector with murine sFas, adsFas, and the control vector adEGFP were constructed using homologous recombination between two plasmids in Escherichia coli. BMT was carried out after the BMMCs were infected with Adenoviruses. The mice models of leukemia/lymphoma were constructed by inoculating female C57BL/6 mice (H-2b) with 10(5) EL4 cells/mouse through caudal vein. Donors of bone marrow grafts were syngeneic male mice. BMMCs were infected with AdsFas or AdEGFP 24 hours before (Group D or E). The following three groups were simultaneously used: Group A, no BMMCs transplanted; Group B, transplanted with BMMCs not infected with adenoviruses; Group C, only transfusing EL4 cells, neither irradiation nor BMT. The hematopoietic reconstitution, generation of leukemia/lymphoma and the survival rate were observed in all groups after BMT. RESULTS: The adenovirus vectors were successfully constructed. The titre of virus after purification was up to 2.5 x 10(11) pfu/ml. Spleen indices examined 11 days after BMT were not obviously different among Group B, D and E (P > 0.05), but indices in Group A were significantly lower than those in the latter three groups (P < 0.01). Counts of leukocytes and platelets on +30 day showed mice were reconstituted satisfactorily in Group B and D, but very low in Group C and E. The Y-chromosomes existed 2 months after BMT and examination of bone marrow cytology showed that Group B and D were almost normal, but Group C and E had plenty of lymphoblast like tumor cells. Tumors were obviously observed in the mice of Group C and E by histopathological examination, but the mice in Group B and D were normal. The survival rates were 0 (0/4) in Group A, 100% in Group B (6/6) and D (16/16), 12.5% (2/16) in Group C and 6.25% (1/16) in Group E respectively. It is demonstrated that, in contrast with the control (Group EGFP), survival rate was significantly increased in the sFas Group (P < 0.01). CONCLUSIONS: The transfer of sFas gene by adenovirus changed the prognosis state of leukemia/lymphoma mice after auto-BMT. The transduction of sFas might block the effect of the immune escape of EL4 cells through FasL. These results could thus provide a new direction to find a way to treat the leukemia and its recurrence after BMT. PMID- 15043785 TI - Adipose tissue-derived stromal cells express neuronal phenotypes. AB - BACKGROUND: Adipose tissue-derived stromal cells (ADSCs) can be greatly expanded in vitro, and induced to differentiate into multiple mesenchymal cell types, including osteogenic, chondrogenic, myogenic, and adipogenic cells. This study was designed to investigate the possibility of ADSCs differentiating into neurons. METHODS: Adipose tissue from rats was digested with collagenase, and adherent stromal cells were cultured. A medium containing a low concentration of fetal bovine serum was adopted to induce the cells to differentiate. ADSCs were identified by immunocytochemistry, and semi-quantitative RT-PCR was applied to detect mRNA expression of neurofilament 1 (NF1), nestin, and neuron-specific enolase (NSE). RESULTS: Nestin-positive cells were found occasionally among ADSCs. ADSCs were found to express NSE mRNA and nestin mRNA, but not NF1 mRNA. ADSCs could differentiate into neuron-like cells in a medium composed of a low concentration of fetal bovine serum, and these differentiated cells displayed complicated neuron-like morphologies. CONCLUSIONS: The data support the hypothesis that adipose tissue contains stem cells capable of differentiating into neurons. These stem cells can overcome their mesenchymal commitment, and may represent an alternative autologous stem cell source for CNS cell transplantation. PMID- 15043786 TI - Co-existence of calcium-binding proteins and gamma-aminobutyric acid or glycine in neurons of the rat medullary dorsal horn. AB - BACKGROUND: We investigated the co-expression of calbindin-D28k (CB), calretinin (CR) and parvalbumin (PV, a combination of the three is referred to as CaBPs) with gamma-aminobutyric acid (GABA) or glycine in neurons of the rat medullary dorsal horn (MDH). METHODS: Immunofluorescence histochemical double-staining for CaBPs and GABA or glycine was performed on the sections from rat MDH. RESULTS: CB , CR-, PV-, GABA- and glycine-like immunoreactive (LI) neurons were differentially observed in all layers of the MDH, but particularly in lamina II. Neurons that exhibited immunoreactivity for both CaBPs and GABA or glycine were also observed mainly in lamina II. A few of them were found in laminae I and III. The percentages of neurons which co-expressed CB/GABA or CB/glycine out of the total numbers of CB- and GABA-LI neurons or CB- and glycine-LI neurons were 5.3% and 12.1% or 4.1% and 10.0%, respectively. The ratios of CR/GABA or CR/glycine co existing neurons out of the total numbers of CR- and GABA-LI neurons or CR- and glycine-LI neurons were 5.8% and 7.6% or 4.4% and 7.1%, respectively. The rates of PV/GABA or PV/glycine co-localized neurons out of the total numbers of PV- and GABA-LI neurons or PV- and glycine-LI neurons were 11.1% and 5.1% or 9.9% and 5.1%, respectively. CONCLUSION: The results indicate that some neurons in the MDH contain both CaBPs and GABA or glycine. PMID- 15043787 TI - Suicide gene therapy of human breast cancer in SCID mice model by the regulation of Tet-On. AB - BACKGROUND: RevTet-On gene expression system was used to deliver the suicide gene tk to human breast cancer cell line MCF-7 and control the tk gene expression level. The animal model of human breast cancer on severe combined immune deficiency (SCID) mice was set up to explore the suicide gene therapy by the regulation of Tet-On. METHODS: Herpes simplex virus-thymidine kinase (HSVtk) gene was inserted into the plasmid pRevTRE and the recombinant retroviral vector pRevTRE/HSVtk was constructed. Using modified calcium phosphate co-precipitation method, two transfections, pRevTRE/HSVtk and pRevTet-On were performed for MCF-7 cell line and selected by hygromycin B and G418. MCF-7 cell line that stably expressed Tet-regulated tk gene was established. HSVtk gene expression in the MCF/TRE/tk/Tet-On cell line was under the control of Doxycycline (Dox). Cell viability was also determined by MTT assay, whereas HSVtk gene expression was analyzed by reverse transcription-PCR (RT-PCR). RESULTS: MCF/TRE/tk/Tet-On cell survival rate was decreased from 100% to less than 20% when ganciclovir (GCV) concentration was increased from 0 to 1000 microg/ml at 1 microg/ml of Dox after 72 hours of GCV administration. At 1 microg/ml of GCV concentration, the cell numbers decreased from 7 x 10(4) cells/ml to 2 x 10(4) cells/ml when Dox concentration was increased from 0 to 1500 ng/ml after 72 hours culture. In addition, bystander effects were generated in vitro when 10% - 25% of transduced MCF-7 cells were mixed in untransduced MCF-7 cells. On the other hand, the human breast cancer models in SCID mice were set up. The tk gene was expressed with the regulated character after MCF/TRE/tk/Tet-On cells were implanted into the female SCID mice 7 days after Dox induction followed by intraperitoneally administration of GCV for 23 days. Subcutaneous tumors in SCID mice that were implanted with MCF/TRE/tk/Tet-On cells shrank remarkably after Dox and GCV administration as compared with the control. CONCLUSION: The human breast tumor cells (MCF-7) expressing HSVtk gene can be eradicated by administration of GCV and induced with tetracycline or its derivative Dox in vitro and in vivo. PMID- 15043789 TI - Telomerase expression in sebaceous carcinoma of the eyelid. AB - BACKGROUND: In humans telomerase is expressed in most cancers and immortal cell lines, and activation of telomerase may play important roles in tumorigenesis and immortalization. This study was to investigate the roles of telomerase activity (TA) and human telomerase RNA (hTR) in sebaceous carcinoma of the eyelid. METHODS: The telomerase repeated amplification protocol (TRAP) was used to demonstrate telomerase activity in 12 cases of sebaceous carcinoma of the eyelid. In situ hybridization (ISH) was used to demonstrate the expression of hTR in 55 cases of paraffin-embedded sebaceous carcinoma of the eyelid, and the results were compared with the proliferative index determined by Mib-1 immuno-labeling, histological patterns and recurrence of the tumor. RESULTS: Different telomerase activity was shown in the 12 cases of sebaceous carcinoma of the eyelid. The positive expression of hTR was 85.5% (47/55) in tumor cells, but not in the adjacent tissues. The positive expression of hTR was correlated with the proliferative activity (as assessed by Mib-1 immunolabelling, r = 0.942, P < 0.001) and the differentiation of sebaceous carcinoma of the eyelid (chi(2) = 17.621, P < 0.001), but not significantly related to tumor recurrence. The level of hTR expression increased with the decrease of differentiation of sebaceous carcinoma of the eyelid. CONCLUSIONS: The results suggest that the up-regulation of telomerase expression plays some roles in tarsal gland carcinogenesis, and the expression of hTR is a useful marker for malignant degree of sebaceous carcinoma of the eyelid. PMID- 15043788 TI - Expression of TN4 gene and its role in human hepatocarcinogenesis from Qidong, a liver cancer risk area. AB - BACKGROUND: We investigated the expression and role of TN4 in the oncogenesis of human hepatocellular carcinoma (HCC) from Qidong which is a HCC risk area. METHODS: The expression of TN4 in HCC was observed using immunohistochemical staining (IHC). TN4 levels were manipulated in human liver cancer cell SMMC7721, using pcDNA3.1 eukaryotic expression constructs designed to express the complete TN4 cDNA. The biological changes of the cells were observed before and after transfection of TN4 and the change of gene expression was analysed by atlas cDNA expression array. RESULTS: Among 100 pairs of samples of HCC, TN4 down-regulation expression and up-regulation expression positive rate were 81% (81/100), 19% (19/100), respectively (P < 0.01). TN4 protein was mainly localized in cytoplasm and membrane. The positive rate of TN4 were 10% (3/30), 100% (70/70) in lymph node metastasis and no lymph node metastasis, respectively (P < 0.01). The growth rates of the derivative SMMC7721-TN4 cell lines were decreased in comparison with that of normal SMMC7721 cells and pcDNA-SMMC7721. Some gene expression was changed before and after transfection of TN4. At 30 days of post-implantation of SMMC7721-TN4, SMMC7721-pcDNA3, SMMC7721 group produced tumors of (301.9 +/- 143.4) mm(3), (2418.7 +/- 362.8) mm(3), (2317.4 +/- 587.8) mm(3), respectively, (P < 0.01). Tumor inhibiting rate was 82.4% in TN4 transfection group. Sections of tumors were observed for their degree of tissue necrosis and there was higher degree of necrosis in tumors of the TN4-SMMC7721 cell group than those of the SMMC7721, SMMC7721-pcDNA groups. CONCLUSIONS: TN4 may play an important role in the oncogenesis of human HCC, especially in Qidong, the HCC risk area and TN4 could be a candidate tumor suppressor gene for HCC. PMID- 15043790 TI - Misoprostol in labour induction of term pregnancy: a meta-analysis. AB - OBJECTIVE: To evaluate the efficacy and safety of misoprostol in term labour induction. DATA SOURCES: Data from published English and Chinese literatures about misoprostol in term labour induction were identified from Medline and CBMdisk (using the search terms "misoprostol" and "labour induction") before 2001; hand searches of reference lists of original studies and reviews (including meta-analyses) and contact with investigators in this field before 2001. STUDY SELECTION: Studies were included if they had data on misoprostol and labour induction. Altogether 623 articles were found and 124 were admitted, including 19,287 cases. DATA EXTRACTION: Data were collected on efficacy and incidence of side-effects of misoprostol and oxytocin. Data were checked for consistency within the published articles and converted into a standard format for incorporation into a central database. DATA SYNTHESIS: The average successful induction rate, rates of caesarean section; incidence of tachysystole, hypertonus of uterus and precipitous labour, and rates of meconium stained amniotic fluid between the misoprostol and oxytocin groups were significantly different (P < 0.05). There were no significant differences between the two groups concerning the average interval from the administration of misoprostol and oxytocin to the onset of labour, duration of the total stage of labour, incidence rate of foetal distress, neonatal asphyxia (1-minute Apgar score < and= 7), postpartum haemorrhage or amount of blood loss in postpartum. CONCLUSIONS: Misoprostol is a superior agent over oxytocin on the induction of term labour, but its application might increase the risk of precipitous labour, abnormal uterine contractions or meconium stained amniotic fluid. Therefore, the dosages and regimens of the agent need further investigation. PMID- 15043791 TI - HMBA-induced differentiation of myeloid leukemic cell lines is associated with altered G1 cell cycle regulators and related genes. PMID- 15043792 TI - A genome-wide allelotype study of primary and corresponding recurrent glioblastoma multiforme in one patient. PMID- 15043793 TI - Relationship of acute left main coronary artery occlusion and ST-segment elevation in lead aVR. PMID- 15043794 TI - Comparative research between Bai-hua Qian-hu, a Chinese traditional plant, and its active ingredient on nuclear factor-kappa and tumor necrosis factor-alpha in isolated ischemia-reperfusion heart of rat. PMID- 15043795 TI - Schizophrenia susceptibility genes on chromosome 13q32. PMID- 15043796 TI - Selective percutaneous radiofrequency thermocoagulation in the treatment of trigeminal neuralgia: report on 1860 cases. PMID- 15043797 TI - Personal computer aided cerebral perfusion imaging with dynamic CT. PMID- 15043799 TI - Dysphagia induced by a cervical osteophyte: a case report of cervical spondylosis. PMID- 15043798 TI - Clinical application of echocardiography in detection of foetal arrhythmia: a retrospective study of 451 foetuses. PMID- 15043800 TI - Hemifacial Spasm. AB - Hemifacial spasm (HFS) is a peripheral movement disorder caused by direct or indirect compression or distortion of the root exit zone of the seventh cranial nerve, which is most commonly compressed by an arterial loop, but also may be compressed by a tumor, cyst, or aneurysm. All patients with HFS should undergo magnetic resonance imaging, with particular attention to the seventh cranial nerve. For patients with HFS who want treatment, there are three options. Oral medications, particularly anticonvulsants, may be useful, but the response rate is low and evidence is almost exclusively anecdotal. Local injection of botulinum toxin into the overactive muscles has a very high rate of success and virtually no serious side effects. Backed by controlled clinical trials, the authors consider it the treatment of choice. Microvascular surgical decompression has the advantage of being potentially curative, and obviates the need for chronic injections with botulinum toxin. However, surgery carries much greater risk than botulinum toxin and the spasm may recur. It is important that surgery is carried out by an experienced neurosurgeon to reduce the risk. PMID- 15043801 TI - Psychosis in Parkinson's Disease. AB - Psychotic symptoms are common and can be a major therapeutic challenge in patients with Parkinson's disease (PD). PD-related psychosis is usually characterized by visual hallucinations or delusions and is most often induced by antiparkinsonian medications. However, other medical conditions, psychoactive medications, sleep disturbances, mood disorders, and cognitive impairments are relevant risk factors. Patients with PD should be continually monitored for factors that can trigger the development of psychotic symptoms, including minor symptoms. This includes ongoing critical re-evaluation of the therapeutic regimen, with adjustments as indicated to optimize function across motor, cognitive, and psychiatric domains. Treatment strategies to reduce psychotic symptoms are determined by the clinical picture. "Benign" symptoms may require only education and reassurance. Antipsychotic medications are required for disabling symptoms and emergency hospitalization may be required for agitation that affects the safety of the patient or others. Medication management is often complex and includes elimination or reduction of antiparkinsonian agents (although this can compromise motor function), management of medical comorbidities, and use of atypical antipsychotics. Clozapine and quetiapine are regarded as the most safe and effective atypical neuroleptics in PD patients. Cholinesterase inhibitors can enhance cognition and may reduce psychotic symptoms. PMID- 15043802 TI - Pain in Parkinson's Disease. AB - Pain is reported by nearly 50% of patients with Parkinson's disease. In some patients, it can be more debilitating than the motor deficits. In order to identify the appropriate treatment strategy for each patient, it is useful to categorize pain syndromes as follows: 1) low DOPA (end of dose wearing off, diphasic, or early morning) painful states are associated with inadequate levels of dopamine receptor stimulation; 2) high DOPA (peak dose) painful states occur at times of maximum levodopa efficacy; and 3) many patients report pain that has no obvious relation to dopaminergic medications or may even be caused by other conditions. Low DOPA painful states are best treated by trying to provide more continuous dopaminergic stimulation and thereby reduce or prevent the number and duration of "off" periods. Adding or increasing the dose of direct-acting dopamine receptor agonists or of catechol-o-methyl transferase inhibitors is the best first-line strategy. Other approaches include increasing the frequency of immediate-release levodopa preparations or using controlled-release preparations. More invasive approaches should be considered only when simpler methods fail. These include deep brain stimulation to the pallidum or the subthalamic nucleus, or direct duodenal continuous infusion of levodopa in patients who are unable to undergo surgery. Pain associated with excessive dopaminergic stimulation usually is a result of dystonia or severe chorea. Reduction of levodopa is the first step in attempting to diminish high DOPA states, followed by reduction or cessation of other dopaminergic agents such as selegiline, catechol-o-methyl transferase inhibitors, or direct-acting dopamine receptor agonists. Adding amantadine can reduce chorea significantly and it should be tried if the potential and actual side effects are tolerable to the patient. Deep brain stimulation is a good final option if medication adjustments are ineffective. Nonspecific pains of Parkinson's disease can be difficult to treat. The effective use of central pain suppressant or analgesics is anecdotal and difficult to verify. In untreated early disease, generalized pain or pain related to joint or muscle immobility may be reduced by effective treatment of the underlying Parkinson's disease. PMID- 15043803 TI - Dementia in Parkinson's Disease. AB - One of the more recently recognized problems in treatment of patients with Parkinson's disease (PD) is development of cognitive dysfunction and, in many cases, frank dementia. As patients with PD live longer, because of improved care and treatment of motor symptoms, dementia in PD is becoming a major contributor to morbidity in the illness. Prevalence studies suggest that up to 30% of patients with PD develop dementia. Dementia in PD patients is often a multifactorial condition. Neuropathologic changes caused by PD itself may cause memory loss. However, some patients with PD and memory decline also have pathologic changes that are more consistent with Alzheimer's disease. Many PD patients have a mix of the two types of pathology. Other factors, such as underlying illnesses, medication side effects, and interaction of therapeutic agents, may contribute to cognitive changes in PD patients. Predictors of development of dementia in PD include advancing age and severity of neurologic symptoms, which may interact with one another to produce this effect. Recent work suggests that tobacco use also may increase risk of PD dementia, despite its possible protective effect against development of PD itself. Presence of psychiatric illness, especially depression, may interfere with cognition and exacerbate memory loss. Reduction in the dose of dopaminergic agents and of other medications may be helpful in partially improving cognitive function in some cases. The balance between improvement of motor function and preservation of cognitive abilities must be weighed, and it is important for clinicians to discuss this trade-off with patients and their families. At this time, there is no US Food and Drug Administration-approved pharmacologic treatment for dementia in PD. However, medication used to treat Alzheimer's disease, such as acetylcholinesterase inhibitors, may slow progression of memory loss in some PD patients. Based on work from small double-blind studies, open-label trials, and case reports, cholinesterase inhibitors may be tried for treatment of dementia in PD, as long as the patient and caregivers understand that these agents are being used on an off-label basis. Surgical intervention, such as deep brain stimulation of the subthalamic nucleus or globus pallidus internus, although useful for treatment of motor symptoms in some PD patients, does not improve cognitive function in most cases and may actually worsen cognition in patients with pre existing dementia. There is no specific exercise regimen or dietary guidelines for patients with PD who develop dementia. However, patients should be encouraged to lead a healthy lifestyle; this may improve overall well-being, which could impact positively on cognition function. Similarly, although assistive devices have not been developed for people with PD who have memory loss, any aid that increases mobility will probably improve mental and physical function. Clinicians should be mindful of the increased caregiver burden posed by PD patients who also have dementia. They should intervene appropriately to prevent caregiver distress and "burn out." Herbal and nutritional supplements have not been shown in clinical trials to be beneficial for treatment of any type of dementia, and thus are not recommended for PD patients with cognitive decline. PMID- 15043804 TI - Restless Legs Syndrome. AB - In the past 10 years, restless legs syndrome (RLS) has gained recognition as a common sleep disorder. There are several therapeutic options in treating patients with RLS. RLS causes significant sleep disturbance and negatively impacts on patient quality of life. Pharmacologic treatment can result in improved sleep and quality of life issues. RLS patients should be evaluated for iron deficiency anemia; iron replacement in deficient patients may lead to a resolution of symptoms or may reduce the severity of their symptoms. For patients with daily symptoms, the initial therapy is dopamine agonists. Low doses given in the evening or 2 hours before bed provide adequate relief of symptoms for many RLS patients. Augmentation can be seen with all dopamine agents, but is most prevalent with levodopa. Levodopa therapy is best used for milder intermittent symptoms or in aggravating situations, such as long car rides. Opiates and antiepileptics remain a beneficial therapy for RLS and are useful in patients who experience pain as part of their RLS. Newer anticonvulsants may provide additional treatment options, but they have yet to undergo clinical trials. Intravenous iron also may provide relief of RLS symptoms; however, dosing and safety issues have not been fully evaluated in a RLS population. PMID- 15043805 TI - Using New Antiepileptic Drugs As Monotherapy. AB - Since 1993, with the advent of felbamate, physicians have been blessed with the addition of eight new antiepileptic medications for the treatment of epilepsy. Increasing evidence is accumulating that these drugs are effective as monotherapy agents. The use of monotherapy minimizes side effects, and newer drugs have fewer interactions than many of the older antiepileptic medications with comparable efficacy. Using the newer antiepileptic agents in monotherapy, even as an initial therapy would appear to be an appropriate clinical decision. PMID- 15043806 TI - Idiopathic Generalized Epilepsy. AB - Successful treatment of idiopathic generalized epilepsy begins with accurate seizure classification. Seizure types, such as absence, myoclonic, and primary generalized tonic-clonic seizures (PGTCS), often can be classified based on a detailed history and inter-ictal electroencephalogram (EEG). Ideally, patients can be classified into specific epilepsy syndromes, such as childhood absence epilepsy, juvenile myoclonic epilepsy (JME), or generalized tonic-clonic seizures on awakening. Idiopathic generalized epilepsy should be distinguished from focal epilepsy with rapid secondary generalization. If this distinction is not clear after history, physical examination, and routine inter-ictal EEG, then ambulatory EEG, video EEG monitoring, or neuroimaging studies may be needed. Ethosuximide, valproate, or lamotrigine are all appropriate first-line choices in the treatment of childhood absence epilepsy. The specific medication should be chosen based on the side effect profiles, dosing formulations, and titration schedules of the medications. The available evidence best supports valproate as the first-line choice in the treatment of JME, although lamotrigine and topiramate may be appropriate choices in this setting. More data specific to JME are needed to clarify the role of medications such as levetiracetam and zonisamide in the treatment of JME. The available evidence to guide the treatment of PGTCS is limited, because most trials did not rigorously exclude patients with focal epilepsy with rapid secondary generalization. Available evidence suggests that valproate is an appropriate first-line choice for PGTCE. Lamotrigine or topiramate also may be appropriate choices. More data are needed to clarify the role of levetiracetam and zonisamide in the treatment of PGCTS. If it is unclear whether a patient has idiopathic generalized epilepsy or focal epilepsy with secondary generalization, then a broad-spectrum anticonvulsant, including valproate, lamotrigine, or topiramate, should be considered. More data are needed to support the broad-spectrum efficacy of levetiracetam and zonisamide. PMID- 15043807 TI - Single Unprovoked Seizures. AB - Unprovoked seizures are common, affecting approximately 4% of the population by age 80. Only approximately 30% to 40% of patients with a first seizure will have a second unprovoked seizure (ie, epilepsy). Treatment with antiepileptic drugs (AEDs) should not be initiated unless the diagnosis of a seizure is firm. Decisions regarding treatment of single unprovoked seizures must balance seizure recurrence risk, the potential impact of a recurrent seizure, the likelihood of adverse effects of treatment, and patient preference. Risk factors for seizure recurrence include a history of remote neurologic insult, epileptiform abnormalities on electroencephalogram, focal structural lesion on neuroimaging, and family history of epilepsy. Adult patients with these risk factors have a recurrence risk of 60% to 70% and usually should be treated with an AED to prevent seizure recurrence. Without risk factors, the recurrence risk is 20% to 30%, and treatment depends on individual risk-to-benefit ratios and patient preference. Treatment of a first unprovoked seizure is often not necessary in childhood, especially if the seizure is part of a benign self-limited syndrome, such as benign Rolandic epilepsy of childhood. Treatment with an AED reduces the risk of seizure recurrence after a single unprovoked seizure. This must be balanced against the risk of adverse effects of AEDs. Treatment of the first seizure does not appear to affect the long-term prognosis of epilepsy. The choice of an AED should be guided by the seizure type and likely epilepsy syndrome diagnosis. Monotherapy is preferable. Standard AED options include phenytoin, carbamazepine, valproate, and phenobarbital. The newer AED, including gabapentin, lamotrigine, topiramate, oxcarbazepine, levetiracetam, and zonisamide, have good efficacy, favorable pharmacokinetic profiles, and often fewer adverse effects, supporting their use early in treatment. Not all of the newer AEDs are approved for use as monotherapy. Patients with single seizures should be counseled about seizure first aid and general safety measures, including precautions regarding swimming alone, engaging in high-risk activities, driving, possible seizure precipitation by photic stimuli (in generalized epilepsy), sleep deprivation, and alcohol. PMID- 15043808 TI - Surgical Treatment for Extratemporal Epilepsy. AB - Partial seizures of extratemporal origin may present unique challenges in the patient with medically refractory seizures. The efficacy of an extratemporal focal cortical resection may be less effective than an anterior temporal lobectomy for intractable epilepsy. The potential operative complications may be increased in individuals with extratemporal epilepsy because of functional cerebral cortex involvement and the need for a large cortical resection to significantly reduce seizure tendency. Partial seizures of extratemporal origin are predominantly associated with frontal lobe epilepsy. The most effective treatment for intractable partial epilepsy is a focal cortical resection with excision of the epileptogenic zone, that is, an area of ictal onset and initial seizure propagation. The preoperative evaluation and operative strategy in patients with partial epilepsy of extratemporal origin associated with pharmacoresistant seizures is determined by the anatomic localization of the epileptogenic zone and the presence of a substrate-directed disorder. The goals of surgical treatment in extratemporal epilepsy include rendering the patient seizure-free, avoiding operative morbidity, and allowing the individual to become a participating and productive member of society. Before surgical treatment, the individual with extratemporal epilepsy will require a comprehensive preoperative evaluation, including routine electroencephalogram (EEG), long-term EEG monitoring, neuropsychologic studies, and magnetic resonance imaging (MRI). Patients with a normal MRI study, conflicting preoperative evaluation, or involvement of suspected functional cerebral cortex would require chronic intracranial EEG monitoring. The rationale for intracranial EEG includes localization of the ictal onset zone or intraoperative functional mapping, or both. Two-fluorodeoxyglucose positron emission tomography studies are usually unremarkable in patients with extratemporal epilepsy and normal MRI scans. Subtraction ictal single photon emission computed tomography coregistered to MRI (SISCOM) study may be useful to demonstrate a localized cerebral perfusion alteration in patients with intractable partial epilepsy. The diagnostic yield of SISCOM has been confirmed in patients with extratemporal epilepsy undergoing surgical treatment. The results of the SISCOM study may tailor the placement of intracranial EEG electrodes and affect the operative strategy. Patients with extratemporal epilepsy overall are less favorable operative candidates than individuals with medial temporal lobe epilepsy. However, individuals with MRI identified lesional pathology of SISCOM-identified perfusion alterations concordant with the epileptogenic zone may be considered for surgical treatment. Chronic intracranial EEG monitoring may be necessary to confirm the localization of the ictal onset zone before epilepsy surgery. Patients with normal neuroimaging studies and extratemporal epilepsy are unlikely to be rendered seizure-free with focal cortical resection and should be considered candidates for other alternative forms of treatment for intractable partial epilepsy. Patients with non-substrate-directed extratemporal epilepsy should undergo a preoperative evaluation and surgical treatment at a comprehensive epilepsy center with extensive experience in chronic intracranial EEG monitoring and contemporary neuroimaging procedures because of the inherently high acuity associated with the operative management clinical disorder. PMID- 15043809 TI - Integration of chemosensory pathways in the Drosophila second-order olfactory centers. AB - BACKGROUND: Behavioral responses to odorants require neurons of the higher olfactory centers to integrate signals detected by different chemosensory neurons. Recent studies revealed stereotypic arborizations of second-order olfactory neurons from the primary olfactory center to the secondary centers, but how third-order neurons read this odor map remained unknown. RESULTS: Using the Drosophila brain as a model system, we analyzed the connectivity patterns between second-order and third-order olfactory neurons. We first isolated three common projection zones in the two secondary centers, the mushroom body (MB) and the lateral horn (LH). Each zone receives converged information via second-order neurons from particular subgroups of antennal-lobe glomeruli. In the MB, third order neurons extend their dendrites across various combinations of these zones, and axons of this heterogeneous population of neurons converge in the output region of the MB. In contrast, arborizations of the third-order neurons in the LH are constrained within a zone. Moreover, different zones of the LH are linked with different brain areas and form preferential associations between distinct subsets of antennal-lobe glomeruli and higher brain regions. CONCLUSIONS: MB is known to be an indispensable site for olfactory learning and memory, whereas LH function is reported to be sufficient for mediating direct nonassociative responses to odors. The structural organization of second-order and third-order neurons suggests that MB is capable of integrating a wide range of odorant information across glomeruli, whereas relatively little integration between different subsets of the olfactory signal repertoire is likely to occur in the LH. PMID- 15043810 TI - A complex of Armadillo, Legless, and Pygopus coactivates dTCF to activate wingless target genes. AB - BACKGROUND: Upon receiving a Wnt signal, cells accumulate beta-catenin (Armadillo in Drosophila), which binds directly to TCF transcription factors, leading to the transcription of Wnt target genes. It is generally thought that beta catenin/Armadillo is a transcriptional coactivator when bound to TCF in the nucleus and that this function is mediated by its C terminus. However, recent findings in Drosophila indicated that Armadillo may activate dTCF in the cytoplasm. RESULTS: Here, I reexamine the mechanism of Armadillo's signaling function in light of Legless and Pygopus, two nuclear factors recently discovered to be essential for this function. I show that Armadillo, in order to activate dTCF, must enter the nucleus and form a complex with Legless and Pygopus. The ability of this complex to stimulate TCF-mediated transcription can be altered by linkage of a strong transcriptional activator or repressor to Armadillo. Furthermore, Armadillo is a strong transcriptional activator when fused to the yeast GAL4 DNA binding domain-an activity that depends on regions of the Armadillo repeat domain that mediate binding to Legless and to chromatin modifying and remodeling factors. Finally, linkage of the N-terminal region of Pygopus, but not the C terminus of Armadillo, to dominant-negative dTCF can restore its signaling activity in transgenic flies. CONCLUSIONS: My evidence argues in favor of a revised coactivator factor model in which Armadillo's coactivator function depends on regions within its Armadillo repeat domain to which Legless/Pygopus and other transcriptional coactivators can bind. In contrast, the C terminus of Armadillo plays a less direct role in this function. PMID- 15043811 TI - The replication fork block protein Fob1 functions as a negative regulator of the FEAR network. AB - BACKGROUND: The protein phosphatase Cdc14 is a key regulator of exit from mitosis in budding yeast. Its activation during anaphase is characterized by dissociation from its inhibitor Cfi1/Net1 in the nucleolus and is controlled by two regulatory networks. The Cdc14 early anaphase release (FEAR) network promotes activation of the phosphatase during early anaphase, whereas the mitotic exit network (MEN) activates Cdc14 during late stages of anaphase. RESULTS: Here we investigate how the FEAR network component Spo12 regulates Cdc14 activation. We identify the replication fork block protein Fob1 as a Spo12-interacting factor. Inactivation of FOB1 leads to premature release of Cdc14 from the nucleolus in metaphase arrested cells. Conversely, high levels of FOB1 delay the release of Cdc14 from the nucleolus. Fob1 associates with Cfi1/Net1, and consistent with this observation, we find that the bulk of Cdc14 localizes to the Fob1 binding region within the rDNA repeats. Finally, we show that Spo12 phosphorylation is cell cycle regulated and affects its binding to Fob1. CONCLUSIONS: Fob1 functions as a negative regulator of the FEAR network. We propose that Fob1 helps to prevent the dissociation of Cdc14 from Cfi1/Net1 prior to anaphase and that Spo12 activation during early anaphase promotes the release of Cdc14 from its inhibitor by antagonizing Fob1 function. PMID- 15043812 TI - Multiple classes of MSL binding sites target dosage compensation to the X chromosome of Drosophila. AB - MSL complexes bind hundreds of sites along the single male X chromosome to achieve dosage compensation in Drosophila. Previously, we proposed that approximately 35 "high-affinity" or "chromatin entry" sites (CES) might nucleate spreading of MSL complexes in cis to paint the X chromosome. This was based on analysis of the first characterized sites roX1 and roX2. roX transgenes attract MSL complex to autosomal locations where it can spread long distances into flanking chromatin. roX1 and roX2 also produce noncoding RNA components of the complex. Here we identify a third site from the 18D10 region of the X chromosome. Like roX genes, 18D binds full and partial MSL complexes in vivo and encompasses a male-specific DNase I hypersensitive site (DHS). Unlike roX genes, the 510 bp 18D site is apparently not transcribed and shows high affinity for MSL complex and spreading only as a multimer. While mapping 18D, we discovered MSL binding to X cosmids that do not carry one of the approximately 35 high-affinity sites. Based on additional analyses of chromosomal transpositions, we conclude that spreading in cis from the roX genes or the approximately 35 originally proposed "entry sites" cannot be the sole mechanism for MSL targeting to the X chromosome. PMID- 15043813 TI - A novel action of histone deacetylase inhibitors in a protein aggresome disease model. AB - Protein inclusions are associated with a number of neurodegenerative diseases including amyotrophic lateral sclerosis (ALS). Whether protein aggregates are toxic or beneficial to cells is not known. In ALS animal models, mutant SOD1 forms aggresome-like structures in motor neurons and astrocytes. To better understand the role of protein aggregation in the progression of disease etiology, we performed a screen for small molecules that disrupt aggresome formation in cultured cells. After screening 20,000 compounds, we obtained two groups of compounds that specifically prevented aggresome formation. One group consists mainly of cardiac glycosides and will be the subject of another study. The second group contains two compounds: one is a known histone deacetylase (HDAC) inhibitor, Scriptaid, and the other is a Flavin analog, DPD. Cells treated with these molecules still contained microaggregates, but these microaggregates were not transported to microtubule organizing centers (MTOCs). The defect in transport was linked to modulation of the dynein/dynactin machinery as treatment with Scriptaid or DPD reversed mSOD-induced insolubilization of the dynactin subunits P50 dynamitin and P150(glued). Our findings suggest a connection between HDAC activity and aggresome formation and also lay the groundwork for a direct test of the role of aggresome formation in ALS etiology. PMID- 15043814 TI - Your own action influences how you perceive another person's action. AB - A growing body of neuroimaging and neurophysiology studies has demonstrated the motor system's involvement in the observation of actions, but the functional significance of this is still unclear. One hypothesis suggests that the motor system decodes observed actions. This hypothesis predicts that performing a concurrent action should influence the perception of an observed action. We tested this prediction by asking subjects to judge the weight of a box lifted by an actor while the subject either lifted or passively held a light or heavy box. We found that actively lifting a box altered the perceptual judgment; an observed box was judged to be heavier when subjects were lifting the light box, and it was judged to be lighter when they were lifting the heavy box. This result is surprising because previous studies have found facilitating effects of movement on perceptual judgments and facilitating effects of observed actions on movements, but here we found the opposite. We hypothesize that this effect can be understood in terms of overlapping neural systems for motor control and action understanding if multiple models of possible observed and performed actions are processed. PMID- 15043815 TI - Axonal heparan sulfate proteoglycans regulate the distribution and efficiency of the repellent slit during midline axon guidance. AB - The presentation of secreted axon guidance factors plays a major role in shaping central nervous system (CNS) connectivity. Recent work suggests that heparan sulfate (HS) regulates guidance factor activity; however, the in vivo axon guidance roles of its carrier proteins (heparan sulfate proteoglycans, or HSPGs) are largely unknown. Here we demonstrate through genetic analysis in vivo that the HSPG Syndecan (Sdc) is critical for the fidelity of Slit repellent signaling at the midline of the Drosophila CNS, consistent with the localization of Sdc to CNS axons. sdc mutants exhibit consistent defects in midline axon guidance, plus potent and specific genetic interactions supporting a model in which HSPGs improve the efficiency of Slit localization and/or signaling. To test this hypothesis, we show that Slit distribution is altered in sdc mutants and that Slit and its receptor bind to Sdc. However, when we compare the function of the transmembrane Sdc to a different class of HSPG that localizes to CNS axons (Dallylike), we find functional redundancy, suggesting that these proteoglycans act as spatially specific carriers of common HS structures that enable growth cones to interact with and perceive Slit as it diffuses away from its source at the CNS midline. PMID- 15043816 TI - Exon junction sequences as cryptic splice sites: implications for intron origin. AB - Introns are flanked by a partially conserved coding sequence that forms the immediate exon junction sequence following intron removal from pre-mRNA. Phylogenetic evidence indicates that these sequences have been targeted by numerous intron insertions during evolution, but little is known about this process. Here, we test the prediction that exon junction sequences were functional splice sites that existed in the coding sequence of genes prior to the insertion of introns. To do this, we experimentally identified nine cryptic splice sites within the coding sequence of actin genes from humans, Arabidopsis, and Physarum by inactivating their normal intron splice sites. We found that seven of these cryptic splice sites correspond exactly to the positions of exon junctions in actin genes from other species. Because actin genes are highly conserved, we could conclude that at least seven actin introns are flanked by cryptic splice sites, and from the phylogenetic evidence, we could also conclude that actin introns were inserted into these cryptic splice sites during evolution. Furthermore, our results indicate that these insertion events were dependent upon the splicing machinery. Because most introns are flanked by similar sequences, our results are likely to be of general relevance. PMID- 15043817 TI - Dispersed male networks in western gorillas. AB - Although kin-selection theory has been widely used to explain the tendency of individuals to bias beneficial behaviors towards relatives living within the same social group, less attention has focused on kin-biased interactions between groups. For animal societies in which females emigrate, as is the case for mountain gorillas (Gorilla beringei beringei), encounters between males in different groups often involve aggressive displays that can escalate to physical violence and fatal injuries. However, recent findings on the little-studied western gorilla (Gorilla gorilla) indicate that interactions between social groups occur more frequently than they do in mountain gorillas and are often, although not always, surprisingly nonaggressive. We investigated the pattern of genetic relationships between individuals of different groups and found evidence suggesting a previously unrecognized "dispersed male network" social structure in western gorillas in which the single males leading social groups were usually related to one or more nearby males. We propose that this provides a basis for extra-group, kin-biased behaviors and may explain the reported peaceful intergroup interactions. Furthermore, these results suggest that a patrilocal social structure, in which males remain in their natal region and potentially benefit from kin associations, is a feature unifying African apes and humans. PMID- 15043818 TI - Geographic variation of caste structure among ant populations. AB - Morphologically distinct worker castes of eusocial insects specialize in different tasks. The relative proportions of these castes and their body sizes represent the demography of a colony that is predicted to vary adaptively with environments. Despite strong theoretical foundations, there has been little empirical evidence for the evolution of colony demography in nature. We show that geographically distinct populations of the ant Pheidole morrisi differ in worker caste ratios and worker body sizes in a manner consistent with microevolutionary divergence. We further show that the developmental mechanism for caste determination accounts for the unique pattern of covariation observed in these two traits. Behavioral data reveal that the frequency of different tasks performed by workers changes in a caste-specific manner when caste ratios are altered and demonstrate the importance of the major caste in colony defense. The population-level variation documented here for P. morrisi colonies supports the predictions of adaptive demography theory and illustrates that developmental mechanisms can play a significant role in shaping the evolution of phenotype at the colony level. PMID- 15043819 TI - Two PAR6 proteins become asymmetrically localized during establishment of polarity in mouse oocytes. AB - Meiotic maturation in mammals is characterized by two asymmetric divisions, leading to the formation of two polar bodies and the female gamete. Whereas the mouse oocyte is a polarized cell, molecules implicated in the establishment of this polarity are still unknown. PAR proteins have been demonstrated to play an important role in cell polarity in many cell types, where they control spindle positioning and asymmetric distribution of determinants. Here we show that two PAR6-related proteins have distinct polarized distributions in mouse oocytes. mPARD6a is first localized on the spindle and then accumulates at the pole nearest the cortex during spindle migration. In the absence of microtubules, the chromosomes still migrate to the cortex, and mPARD6a was found associated with the chromosomes and was facing the cortex. mPARD6a is the first identified protein to associate with the spindle during spindle migration and to relocalize to the chromosomes in the absence of microtubule behavior, suggesting a role in spindle migration. The other protein, mPARD6b, was found on spindle microtubules until entry into meiosis II and relocalized to the cortex at the animal pole during metaphase II arrest. mPARD6b is the first identified protein to localize to the animal pole of the mouse oocyte and likely contributes to the polarization of the cortex. PMID- 15043820 TI - A mechanism for sensing noise damage in the inner ear. AB - Our sense of hearing requires functional sensory hair cells. Throughout life those hair cells are subjected to various traumas, the most common being loud sound. The primary effect of acoustic trauma is manifested as damage to the delicate mechanosensory apparatus of the hair cell stereocilia. This may eventually lead to hair cell death and irreversible deafness. Little is known about the way in which noxious sound stimuli affect individual cellular components of the auditory sensory epithelium. However, studies in different types of cell cultures have shown that damage and mechanical stimulation can activate changes in intracellular free calcium concentration ([Ca(2+)](i)) and elicit intercellular Ca(2+) waves. Thus an attractive hypothesis is that changes in [Ca(2+)](i), propagating as a wave through support cells in the organ of Corti, may constitute a fundamental mechanism to signal the occurrence of hair cell damage. The mechanism we describe here exhibits nanomolar sensitivity to extracellular ATP, involves regenerative propagation of intercellular calcium waves due to ATP originating from hair cells, and depends on functional IP(3) sensitive intracellular stores in support cells. PMID- 15043821 TI - Regulation of retinal cone bipolar cell differentiation and photopic vision by the CVC homeobox gene Vsx1. AB - Cone bipolar cells of the vertebrate retina connect photoreceptors with ganglion cells to mediate photopic vision. Despite this important role, the mechanisms that regulate cone bipolar cell differentiation are poorly understood. VSX1 is a CVC domain homeoprotein specifically expressed in cone bipolar cells. To determine the function of VSX1, we generated Vsx1 mutant mice and found that Vsx1 mutant retinal cells form but do not differentiate a mature cone bipolar cell phenotype. Electrophysiological studies demonstrated that Vsx1 mutant mice have defects in their cone visual pathway, whereas the rod visual pathway was unaffected. Thus, Vsx1 is required for cone bipolar cell differentiation and regulates photopic vision perception. PMID- 15043822 TI - Green light flickering for the UK's first GM crop. PMID- 15043823 TI - Red queen running on gene technologies. PMID- 15043825 TI - Hugo J. Bellen. PMID- 15043826 TI - Gang welfare. PMID- 15043827 TI - AMPK. PMID- 15043828 TI - Animal navigation. PMID- 15043829 TI - Hypothesis: a helix of ankyrin repeats of the NOMPC-TRP ion channel is the gating spring of mechanoreceptors. PMID- 15043830 TI - Coronavirus outbreak in cheetahs: lessons for SARS. PMID- 15043832 TI - Inner ear: Ca(2+)n you feel the noise? AB - The death of hair cells in the inner ear as a result of exposure to loud noise can lead to irreversible deafness. New work shows that the mammalian cochlea can sense noxious sounds and use Ca(2+) waves to rapidly propagate hair cell damage signals. PMID- 15043831 TI - Germ cells: finding programs of mass repression. AB - Transcription is globally silenced in the germline of animals. Recent studies have shown that, in Caenorhabditis elegans, this silencing is initially mediated through direct repression, but in Drosophila, the factors involved include pgc, a non-coding cytoplasmic RNA. Why are these mechanisms so diverse and complex? PMID- 15043833 TI - Genome size: does bigger mean worse? AB - Genome sizes vary enormously. This variation in DNA content correlates with effective population size, suggesting that deleterious additions to the genome can accumulate in small populations. On this view, the increased complexity of biological functions associated with large genomes partly reflects evolutionary degeneration. PMID- 15043834 TI - Neuroscience: what you see and hear is what you get. AB - The brain receives signals from a variety of sources; for example, visual and auditory signals can both indicate the direction of a stimulus, but with differing precision. A recent study has shed light on the way that the brain combines these signals to achieve the best estimate possible. PMID- 15043835 TI - Embryonic axes: the long and short of it in the mouse. AB - Contrary to conventional wisdom, recent studies indicate that, during mouse development, the embryo's anterior-posterior axis shifts from the shorter transverse axis to the orthogonal longer one as the shape of the pre-gastrula stage embryo is remodelled. PMID- 15043836 TI - Bacterial shape: concave coiled coils curve caulobacter. AB - Bacterial cells exhibit a wide variety of shapes. Recent results indicate that the characteristic crescent shape of Caulobacter crescentus depends upon an inter mediate filament-like protein that localizes to the concave side of the cell. PMID- 15043837 TI - Evolution: bacterial mutation in stationary phase. AB - A recent study indicates that the genomic mutation rate of the gut bacterium Escherichia coli is substantially higher in nongrowing than growing cultures. These findings are important in the light of the ongoing controversy over the generality and robustness of stationary phase mutagenesis and its evolutionary implications. PMID- 15043838 TI - Sensorimotor gating: startle submits to presynaptic inhibition. AB - Prepulse inhibition is a type of sensorimotor gating that is disrupted in some neuropsychiatric disorders. Its cellular basis has eluded investigators but now, using a molluscan model system, a cellular mechanism has been established. PMID- 15043839 TI - Membrane curvature: how BAR domains bend bilayers. AB - An important new structure suggests the BAR domain is a membrane-binding module that can both produce and sense membrane curvature. BAR resembles a banana that binds membranes electrostatically through its positively charged, concave surface. PMID- 15043840 TI - Silkworm-pheophorbide alpha mediated photodynamic therapy against B16F10 pigmented melanoma. AB - In order to apply photodynamic therapy (PDT) to pigmented melanoma, the efficacy of PDT mediated by pheophorbide alpha from silkworm excreta (SPbalpha) and commercial Photofrin against B16F10 melanoma was comparatively studied from the in vivo assay using C57BL/6J mice. From in vitro PDT assay, the proliferation of B16F10 cells treated with SPbalpha (more than 0.5 microg/ml) and light illumination (1.2 J/cm2) were significantly inhibited with the necrotic response. This indicated that the photocytotoxicity of SPbalpha (665 nm) was not influenced by melanin from melanoma. From the assessment of the in vivo photosensitizing activity, the tumor growth was further delayed in groups treated with SPbalpha/PDT compared to that treated with Photofrin /PDT. The survival rate of tumor bearing mice treated with SPbalpha/PDT was closely associated with its photosensitizing effect. In addition, the photosensitizing effect of SPbalpha/PDT showed a dose dependent tendency in light illumination. These results demonstrated that B16F10 melanoma cells were significantly photosensitized by SPbalpha/PDT, regardless of the influence of melanin from melanoma, and SPbalpha/PDT at very low drug dose (1 mg/kg) and light dose (1.2 J/cm2) showed the photosensitizing efficacy surpassing Photofrin against B16F10 melanoma in mice system. PMID- 15043841 TI - A comparative study of 632.8 and 532 nm laser irradiation on some rheological factors in human blood in vitro. AB - The effects of laser irradiation with 632.8 and 532 nm on rheological properties of blood were comparatively studied in vitro. Under the irradiation condition of 30 mW, laser irradiation of blood samples using a spot diameter of 5 mm with each laser, showed promising results in the modulation of hemorheological properties. When blood samples from patients with abnormally high values of erythrocyte sedimentation rate (ESR) were irradiated, the values of ESR were lowered statistically by either of the 632.8 or 532 nm lasers. The laser irradiation reduced blood viscosities at different shear rates (10-110 S(-1)) for the hyper viscosity blood samples. Laser irradiation increased the electrophoretic mobility (EPM) of erythrocytes when the values of the sample's EPM were abnormally slow. The erythrocyte deformability was enhanced by laser irradiation when the deformability of the sample from the patients was originally poor. For verifying the improvement of laser irradiation on erythrocyte deformability, the typical erythrocyte samples with poor deformability were produced by the pre-treatment of the erythrocytes with Ca(2+). The deformability of these erythrocyte samples was also improved after laser irradiation. These results suggest that membrane-bound hemoglobin (Hbm) might be the initial site of the interaction, since Hbm is the main cause of poor deformability when erythrocytes were treated with Ca(2+). In all experiments including ESR, blood viscosity, EPM and erythrocyte deformability, the 532 nm laser demonstrated more efficient effects on modulating rheological properties than 632.8 nm laser. This wavelength effect is consistent with the absorption spectrum of hemoglobin, reflecting that hemoglobin may be one of the action targets under laser irradiation. PMID- 15043842 TI - Ion transport across model lipid membranes containing light-harvesting complex II: an effect of light. AB - The effect of light on proton transport across lipid membranes of small unilamellar liposomes containing incorporated major light-harvesting pigment protein complex of Photosystem II (LHCII) has been studied with the application of pH-sensitive dyes entrapped inside vesicles. Proton permeability coefficient for LHCII-modified membranes was found to be about twice as high as in the case of the control pure lipid vesicles. Illumination of the samples with light absorbed by the LHCII-bound photosynthetic pigments considerably affects the kinetics of proton transport: it increases the rate and decreases the steady state level of proton gradient across the membranes. The effect was interpreted in terms of heat-induced conformational changes of LHCII molecular structures that affect proton buffering capacity of this protein. Both the control and the LHCII-modified lipid membranes have been found to be practically impermeable to Ca(++) ions, as demonstrated by fluorescence of liposome-entrapped calcium sensitive probe calcium crimson. The slight differences in the proton transport across the LHCII-containing membranes under the presence of Ca(++) suggest calcium binding to this antenna protein. PMID- 15043843 TI - Increased level of oxidative stress in genomically unstable cell clones. AB - Recently, we reported that ultraviolet radiation induces delayed mutations in mammalian cells. At the same level of cell death the oxidative component of sunlight (ultraviolet A radiation) was as potent in inducing this kind of genomic instability as ultraviolet B radiation. Ultraviolet B radiation predominantly harms cells by direct damage to DNA and thus is much more mutagenic than ultraviolet A radiation. From that study, clones with a significantly increased mutation rate in the hypoxanthine phosphoribosyl transferase gene were obtained. These genomically unstable clones were also found to have a higher variance in the number of chromosomes than the unirradiated control cells, indicating chromosomal instability. The mechanisms for induction and maintenance of radiation induced genomic instability are not known, but some studies suggest that reactive oxygen species might be involved. In the present study, we have measured the level of potentially mutagenic peroxides in the genomically unstable clones. The levels of intracellular peroxides and lipid peroxides were measured using the probes dihydrorhodamine 123 and diphenyl-1-pyrenyl-phosphine, respectively. The unstable clones had elevated levels of oxidants, supporting the hypothesis that intermediate reactive oxygen species might have a role in the maintenance of genomic instability induced by ultraviolet radiation. PMID- 15043844 TI - Enhanced photo-stability, thermal-stability and aqueous-stability of indocyanine green in polymeric nanoparticulate systems. AB - Photo-degradation, thermal-degradation and aqueous-instability of indocyanine green (ICG) limits its application as a fluorescence contrast agent for imaging purposes. Thus, the objective of this study is to develop polymeric nanoparticles entrapping ICG and to establish its effectiveness in providing photo-stability, thermal stability and aqueous stability to ICG. Nanoparticles entrapping ICG were engineered, characterized and the degradation kinetics of ICG in the nanoparticles was investigated in aqueous media. The entrapment of ICG in the nanoparticles causes a shift in its wavelength of peak fluorescence and a decrease in its peak fluorescence intensity. The degradation of ICG in aqueous nanoparticle suspension followed first-order kinetics for the time period studied. ICG entrapment in the nanoparticles enhanced aqueous-stability of ICG (half-life, t(1/2) was 72.2+/-6.1 h for ICG in the nanoparticles as compared to 16.8+/-1.5 h for free ICG solution), photo-stability of ICG (t(1/2) was 73.7+/ 7.5 h for ICG in the nanoparticles as compared to 14.4+/-2.4 h for free ICG solution when exposed to room light from two 32 W normal fluorescent tubes) and thermal-stability of ICG (t(1/2) of ICG at 42 degrees C was 62.4+/-1.7 h for ICG in the nanoparticles as compared to 10.1+/-0.6 h for free ICG solution). PMID- 15043845 TI - Binding of wogonin to human serum albumin: a common binding site of wogonin in subdomain IIA. AB - The binding of wogonin to human serum albumin (HSA) has been studied by spectroscopic method including circular dichroism (CD), infrared spectra (IR) and fluorescence spectra. The fluorescence properties of HSA were examined in presence of wogonin and the fluorescence intensity of HSA was significantly decreased in the presence of wogonin. The binding parameters of wogonin were studied from the fluorescence decreasing of HSA by the fluoremetric titrations. The Stern-Volmer plots indicated that the binding of wogonin to HSA at 296, 303, 310 K is characterized by one binding site with the binding constant K(S-V) at 1.872 x 10(5), 1.561 x 10(5), 1.392 x 10(5), respectively, which are good agreement with the results from the Scatchard plots. The binding process was exothermic, enthalpy driven and spontaneous, as indicated by the thermodynamic analyses, and the major part of the binding energy is hydrophobic interaction, which were consistent with the result of molecule modelling study, and there are also a numbers of hydrogen bonds between wogonin and HSA. Furthermore, the displacement experiments indicate that wogonin can bind to the subdomain IIA, that is, the site I of HSA, which is also good agreement with the result of molecule modelling study. PMID- 15043847 TI - Introduction to the state of the art of nutrition support in Japan. PMID- 15043846 TI - Up-regulation by phytochrome A of the active protochlorophyllide, Pchlide655, biosynthesis in dicots under far-red light. AB - It is well-documented that phytochrome A (phyA) down-regulates the synthesis of NADPH:protochlorophyllide (Pchlide) oxidoreductase and active Pchlide(655) under far-red light (FR). In this work, we demonstrate that phyA can up-regulate the synthesis of Pchlide(655) under FR as well and that its sign and extent depend on plant species and tissue. With the use of fluorescence spectroscopy, it was found that [Pchlide(655)] in the upper stems of FR-grown seedlings of pea and tobacco increased > or =10-fold and much lower in cotyledons or leaves as compared with the dark-grown. In the upper stems of Arabidopsis and tomato, the positive effect of FR was low, 1.2- to 1.5-fold, and the negative effect of FR was seen in cotyledons. In stems of wild-type (WT) tobacco and its line overexpressing full length oat phyA (FL), we observed gross stimulating effect of FR while in its line overexpressing N-terminally truncated (Delta7-69) oat phyA (NA) it was low. Because WT and FL comprise both native phyA forms, phyA' and phyA", while NA, only phyA", the regulation under FR can be associated with phyA', while phyA" inhibits the action of phyA'. In etiolated seedlings of the NA line, [Pchlide(655)] was much higher than in those of WT and FL suggesting that phyA" may have relation to this enhancement. The regulation of Pchlide(633) in contrast to Pchlide(655) was positive independent of the plant species and tissue. PMID- 15043848 TI - Dietary calcium, sodium, phosphorus, and protein and bone metabolism in elderly Japanese women: a pilot study using the duplicate portion sampling method. AB - OBJECTIVES: Associations between dietary factors and bone metabolism in Asians have not been thoroughly investigated. The purpose of this study was to determine the effect of dietary intake of calcium (Ca), phosphorus (P), sodium (Na), and protein on bone metabolism in elderly Japanese women by the duplicate portion sampling method. METHODS: The subjects were 43 healthy female volunteers (mean age, 68.3 y; standard deviation, 6.8). Dietary nutrients were directly determined by using a 24-h duplicate meal portion. Serum osteocalcin and bone alkaline phosphatase and urinary deoxypyridinoline (DPD) and type I collagen cross-linked N-telopeptides (NTX-I) were measured as markers of bone turnover. Hormones related to bone metabolism, including serum 25-hydroxyvitamin D(3), 1,25 dihydroxyvitamin D(3), and intact parathyroid hormone also were determined. Urinary parameters were corrected for urinary creatinine concentration. RESULTS: The mean (standard deviation) daily dietary intakes of Ca, P, Na, and protein were 660 (195) mg, 996 (208) mg, 4080 (1142) mg, and 63.9 (15.5) g, respectively. Dietary Ca was negatively correlated with urinary DPD (r = -0.417, P = 0.005) and NTX-I (r = -0.324, P = 0.034), and dietary P was negatively correlated with urinary DPD (r = -0.307, P = 0.045). Multiple regression analysis showed that only dietary Ca was associated with urinary DPD and NTX-I. Dietary Ca intake was significantly associated with bone resorption markers, but no other dietary factors were associated with any of the biochemical markers. CONCLUSION: The major factor having an adverse effect on the bone health of the elderly Japanese women is low Ca intake and not other dietary minerals or protein. PMID- 15043849 TI - Refeeding syndrome in patients with gastrointestinal fistula. AB - OBJECTIVE: Although refeeding syndrome has been well documented in starved patients, obese patients, those with anorexia nervosa, malnourished elderly individuals, and certain postoperative patients, little is known about the presence and the importance of refeeding syndrome in patients with gastrointestinal fistula and insufficient nutrition support over the long term. The objective of this study was to estimate the morbidity of this syndrome in these patients, to assess the safety and efficacy of our graduated refeeding regimen, and to emphasize the importance of this syndrome. METHODS: One hundred fifty-eight patients with gastrointestinal fistula during the past 2 y were reviewed. RESULTS: Fifteen of these patients were diagnosed as having refeeding syndrome. They were started on the refeeding procedure according to our regimen, and changes in their serum levels of electrolytes were recorded. The symptoms and signs they presented were noted. All patients were successfully advanced to full nutrition support. During the refeeding procedure, patients presented with weakness, paralysis of limbs, slight dyspnea, paresthesia, tachycardia, edema, and diarrhea. Serum phosphorus concentration decreased in all patients within 24 h of refeeding, reaching a mean nadir after 3.3 +/- 1.5 d and another 6.1 +/- 2.1 d to return to above 0.70 mM/L upon phosphorus supplementation. Three patients treated with growth hormone presented more severe hypophosphatemia (<0.20 mM/L) than the others. CONCLUSIONS: 1) Refeeding syndrome occurs commonly in patients with malnutrition secondary to gastrointestinal fistula. 2) Alterations in phosphate metabolism are central to the refeeding syndrome. 3) Supplementation with electrolytes (including especially phosphate) and vitamins is the focal point of the treatment of this syndrome. 4) Growth hormone treatment may aggravate hypophosphatemia. PMID- 15043850 TI - Problems in serum albumin measurement and clinical significance of albumin microheterogeneity in cirrhotics. AB - OBJECTIVES: To clarify problems with the determination of serum albumin levels, the definition of hypoalbuminemia, and the implications of microheterogeneity of albumin, serum albumin was measured by using dye-binding methods and the authentic method (immunoassay) in patients with liver cirrhosis and healthy subjects. METHODS: We enrolled 103 patients with liver cirrhosis and 36 healthy subjects. Serum albumin levels were analyzed by immunoassay and the bromcresol green and bromcresol purple methods. Oxidized albumin and glycoalbumin were determined by high-performance liquid chromatography. RESULTS: In cirrhotic patients, serum albumin levels measured by the bromcresol green method was about 0.2 g/dL higher than that by immunoassay. Serum albumin levels measured by the bromcresol purple method also was higher in cirrhotic patients than those measured by immunoassay and varied widely. In addition, extensive variation was found across serum albumin levels determined by the bromcresol green method at individual institutions (five university hospitals) and those determined by immunoassay at a contract laboratory. The percentages of oxidized albumin and glycoalbumin within total serum albumin increased with progression of liver disease. Further, an increase in oxidized albumin led to an increase in the albumin level as measured by the bromcresol purple method. CONCLUSION: These results show that adequate assessment of the pathophysiology and prognosis of patients with liver cirrhosis and the efficacy of treatment is not possible with dye-binding methods for determination of serum albumin. Further, the conventional definition of hypoalbuminemia as a serum albumin level of 3.5 g/dL or lower should be reconsidered, and the clinical implications of qualitative changes in albumin should be investigated in consideration of the microheterogeneity of albumin, such as oxidized albumin and glycoalbumin. PMID- 15043851 TI - Vitamin E supplementation increases polyunsaturated fatty acids of RBC membrane in HCV-infected patients. AB - OBJECTIVE: We investigated the effects of vitamin E supplementation on the fatty acid composition of red blood cell membrane phospholipids and on the clinical observations in patients with hepatitis C virus. METHOD: Eight patients and control subjects were administered 500 mg/d of d-alpha-tocopherol for 12 wk. The alpha-tocopherol and fatty acid composition of phospholipids in red blood cells were analyzed before, at 4, 8, and 12 wk, and after 4 wk of washout of vitamin E administration. RESULTS: The alpha-tocopherol concentration in red blood cells increased 2.37-fold of the basal level during vitamin E supplementation. Serum alanine aminotransferase levels increased in five of eight patients with vitamin E supplementation. The arachidonic acid level, docosahexaenoic acid level, and ratio of polyunsaturated to saturated fatty acid in red blood cell membrane phospholipids, which were significantly lower in the patients than in the control subjects, were elevated at 8 and 12 wk after vitamin E supplementation. The improvement in fatty acid composition was observed particularly in the patients who responded to the vitamin E therapy. CONCLUSIONS: Vitamin E therapy for the prevention of disease progression in patients with hepatitis C virus may be effective. PMID- 15043852 TI - Immunonutrition risk factors of respiratory complications after esophagectomy. AB - BACKGROUND: There are many reports suggesting predictive factors for respiratory complications after esophagectomy, but few studies have focused on this problem from the aspect of immunonutrition. METHODS: A series of 45 esophageal cancer patients who underwent potentially curative resection between April 1996 and March 2001 was enrolled in this study. Preoperative and intraoperative variables were analyzed for a correlation between risk factors and respiratory complications. In this study, immunonutrition variables as assessed by ultrasonography and indirect calorimetry were used. Uni- and multivariate analyses were performed to determine the predictive factors. RESULTS: Of 45 patients, nine patients (20%) developed respiratory complications. In univariate analysis, forced expiratory volume in 1 s per body surface area (m(2)) in preoperative respiratory parameters, respiratory quotient, and caloric contributions of fat (percentage) and carbohydrate according to indirect calorimetry, and serum CH50 level significantly influenced the occurrence of postoperative respiratory complications. In multivariate analysis, the caloric contribution of fat (percentage) by indirect calorimetry and serum alpha(1) antitrypsin (serine protease inhibitor) independently affected the occurrence of respiratory complications. CONCLUSIONS: Preoperative excessive fat oxidation (potential starvation) and alpha(1)-antitrypsin were independent predictive factors for postoperative complications. Nutrition support such as enteral feeding to improve this malnourished state would reduce the incidence of postoperative respiratory complications. PMID- 15043853 TI - Efficacy of long-term dietary restriction of total calories, fat, iron, and protein in patients with chronic hepatitis C virus. AB - OBJECTIVES: A diet restrictive in total calories, fat, iron, and protein intake reduces serum alanine aminotransferase levels in patients with long-term hepatitis C virus infection. However, whether long-term dietary therapy causes adverse effects such as malnutrition and anemia due to a shortage of energy intake is not clear. We evaluated the balance of energy intake and changes in physical and hematologic indices of nutrition after a long-term dietary therapy. METHODS: Twenty-two patients with long-term hepatitis C virus infection that did not respond to or who were able or unwilling to take interferon therapy were enrolled in this study. Our prescriptions included 7 mg/d or less of iron, 30 kcal. kg(-1). d(-1) of energy, 1.1 to 1.2 g. kg(-1). d(-1) of protein, and a fat energy fraction of 20%. Patients were followed for 24 mo. RESULTS: Mean body fat percentage was 24.6% at entry and was significantly reduced after the diet prescription. Mean serum ferritin decreased significantly from 376 ng/mL at entry to 141 ng/mL after 24 mo. Mean serum alanine aminotransferase levels decreased significantly from 66 to 49 IU/L. Mean levels of hemoglobin, serum albumin, and cholinesterase did not change significantly during the follow-up period. CONCLUSIONS: These results suggest that restriction of energy, fat, iron, and protein intakes is safely tolerated, so its long-term use should be recommended to patients with long-term infection with hepatitis C virus. PMID- 15043854 TI - Translocation of Salmonella typhimurium in rats on total parenteral nutrition correlates with changes in intestinal morphology and mucus gel. AB - OBJECTIVE: We tested whether alterations in intestinal morphology and mucus gel correlate with differences in Salmonella typhimurium translocation between rats treated with total parenteral nutrition (TPN) and rats given a diet of chow. METHODS: Twenty-seven male Wistar rats were assigned to one of two groups: one received TPN for 14 d and the other (control) received standard rat chow and water ad libitum. Salmonella typhimurium (5 x 10(8) cells; GIFU 12142) was injected into a closed ileal loop. Portal venous blood (PVB), inferior vena cava blood (IVCB), and mesenteric lymph nodes (MLNs) were sampled for evaluation of bacterial translocation. Sections of the loop were prepared and stained with hematoxylin and eosin, periodic acid-Schiff (PAS), and fluorescein isothiocyanate labeled Ulex europaeus agglutinin I (FITC-UEA-I) for image analysis. Perimeter, mucosal thickness, villus area, and positively stained mucus area were measured. A fluorescent antibody study was also done. RESULTS: Organisms were found in cultures of 1 in 13 control rats and 9 in 14 TPN rats. There were more bacteria in MLNs than in PVB or IVCB. There was no increase in the number of bacteria over time in PVB, IVCB, or MLNs. Perimeter and villus area (P < 0.001) and mucosal thickness (P < 0.01) were significantly smaller in the TPN group than in the control group. The positively stained mucus area was significantly smaller in the TPN group than in the control group (P < 0.05 with PAS, P < 0.01 with FITC-UEA I). Salmonella typhimurium invaded specifically through Peyer's patches. In all culture-negative samples, bacteria were trapped by the mucous layer, with a very small number attached to the epithelial surface. CONCLUSION: Significant villous atrophy and reduction of mucus play an important role in the rapid translocation of S. typhimurium through Peyer's patches in rats after 2 wk of TPN. PMID- 15043855 TI - Viscous properties of pig cecal contents and the contribution of solid particles to viscosity. AB - OBJECTIVE: It is believed that solid particles are not responsible for digesta viscosity. However, solid particles are responsible for the viscosity of whole blood and fiber suspension such as wood pulp. Therefore, we measured viscosity of gut contents with and without solid particles. We also determined the contribution of solid particles to viscoelastic characters of gut contents. METHODS: We measured the coefficient of viscosity, shear rate, and shear stress of pig cecal contents by using a tube-flow viscometer before and after removal of particles. RESULTS: The coefficient of viscosity of intact pig cecal contents was negatively correlated with shear rate, a typical character of non-Newtonian fluids. The removal of solid particles from the contents resulted in a Newtonian fluid with very low viscosity. CONCLUSION: Solid particles were mainly responsible for the viscoelastic character of pig cecal contents. Accordingly, the contribution of indigestible solid food components, sloughed epithelial cells, and bacterial cell bodies to the viscosity of gut contents should be re evaluated. PMID- 15043856 TI - Dietary nucleosides and nucleotides do not affect tumor incidence but reduce amyloidosis incidence in B6C3F1 mice irradiated with californium-252. AB - OBJECTIVE: We investigated the effects of a dietary mixture of nucleosides and nucleotides (NS) on the systemic incidence rates of postirradiation carcinogenesis and non-neoplastic lesions in mice. METHODS: Five-week-old male B6C3F1 mice were fed AIN-76B Purified Diet supplemented with NS for 1 wk and 13 mo before and after irradiation of neutron with californium-252 ((252)Cf); specifically NS was added to the AIN-76B Purified Diet (without nucleotide) to obtain a final concentration of 0%, 0.5%, or 2.5% NS. A commercial stock diet was also given to mice, and half of the mice were irradiated. Both irradiated and non irradiated mice were used for reference controls. RESULTS: The incidence of liver tumors in each NS group was lower than that in the reference control group (P < 0.01), but there were no differences between the 0%, 0.5%, and 2.5% NS groups. In contrast, the incidence rate of mice with non-neoplastic lesions in the 0% NS group was significantly higher than the reference control group (P < 0.05). This higher incidence of mice with non-neoplastic lesions was significantly decreased upon supplementation of the nucleotide-free diet with 0.5% or 2.5% NS (P < 0.01 and P < 0.05, respectively). Of the non-neoplastic lesions observed, the incidence of amyloidosis was decreased significantly upon supplementation of the nucleotide-free diet with 0.5% NS (P < 0.05). CONCLUSION: Supplementation of a nucleotide-free diet with NS inhibits the development of non-neoplastic lesions, such as those associated with amyloidosis, without promoting the carcinogenesis induced by (252)Cf irradiation. PMID- 15043857 TI - Gamma-linolenic acid prevents conjugated linoleic acid-induced fatty liver in mice. AB - OBJECTIVE: The present study was done to clarify the mechanism by which conjugated linoleic acid (CLA) induces fatty liver in mice and to attenuate this symptom by adding other dietary fatty acids. METHODS: Mice were given CLA short (12 h) or long (4 wk) term or given CLA with linoleic acid (LA) or gamma linolenic acid (GLA) in the long term (4 wk). Total lipids, triacylglycerol, and prostaglandin E(2) (PGE(2)) levels in the liver were determined. RESULTS: A single administration of CLA significantly increased PGE(2) levels in the liver 12 h after administration. However, long-term administration of CLA significantly decreased the liver PGE(2) level and induced fatty liver. GLA increased PGE(2) levels, and coadministration with GLA, but not with LA, prevented the CLA-induced fatty liver. CONCLUSIONS: These data suggest that CLA initially stimulates PGE(2) production followed by depletion of PGE(2) sources in the liver. The fatty liver associated with PGE(2) reduction by CLA ingestion can be attenuated by GLA in mice. PMID- 15043858 TI - Modification of concanavalin A-dependent proliferation by phosphatidylcholines isolated from deer antler, Cervus elaphus. AB - OBJECTIVE: The immunomodulatory effect of deer antler, which is used as traditional medicine, has been known, but the active component of antlers from Cervus elaphus has not been identified. In this study, we identified the immunomodulator from C. elaphus and examined its biological activities on the immune system. METHODS: To identify an immunomodulator, we used bioassay-guided fractionation after silica gel column chromatography. Structural analysis was performed with one- and two-dimensional nuclear magnetic resonance techniques and tandem mass spectrometry coupled with fast atom bombardment. RESULTS: The subfraction, phosphatidylcholines, isolated 70% ethanol extract of C. elaphus induced the proliferation of spleen cells in synergy with concanavalin A. According to the structural analysis, phosphatidylcholines were classified as a family (1,2-alkyl-sn-glycerol-3-phosphocholines) containing arachidonyl (C20:4), stearoyl (C18:0), oleoyl (C18:1), linoleoyl (C18:2), palmitoyl (C16:0), and myristoyl (C14:0) chains in their fatty acyl chains. Because the unsaturated fatty acids showed an inhibitory effect on the immune system, dialkyl phosphatidylcholines with different chain lengths from C10:0 to C20:0 that stimulate the proliferation of spleen cells were examined extensively. Among other saturated phosphatidylcholines used, dimyristoyl phosphatidylcholine (C14:0) induced the proliferation of spleen cells more efficiently, whereas dimyristoleoyl phosphatidylcholine (C14:1) effected little change in the proliferation of spleen cells. CONCLUSIONS: These data collectively suggest that phosphatidylcholines with saturated fatty acyl chains are immunostimulating factors. They may modify the proliferation of known mitogens. Further, chain length and saturation of the fatty acids may play important roles in the proliferation of spleen cells. PMID- 15043859 TI - The ability of biofilm formation does not influence virulence of Staphylococcus aureus and host response in a mouse tissue cage infection model. AB - The virulence of Staphylococcus aureus Sa113 (SA113) and an isogenic ica deletion mutant (ica-), deficient in the production of polysaccharide intercellular adhesin (PIA), which is crucial for biofilm formation, was compared in a mouse tissue cage infection model. The minimal infective doses for the induction of persistent tissue infections in C57BL/6 mice were 10(3) CFU for both SA113 and the ica- mutant. Bacterial growth, initial adherence to surfaces within the implants and the course of inflammation including growth-dependent host TNF and MIP-2 release, influx of phagocytes and an accumulation of dead leukocytes were similar as well. Since SA113 expressed PIA in vivo, we could demonstrate that PIA and the lack of biofilm formation did not influence the capacity of S. aureus to induce persistent infections and did not modulate host responses in the mouse tissue cage model. PMID- 15043860 TI - Identification of a nuclear targeting signal in YopM from Yersinia spp. AB - YopM is a type III secretion effector from Yersinia which contributes to pathogenicity but whose action still remains unclear. It is an acidic, leucine rich repeats (LRR) containing protein which migrates to the nucleus of target cells in spite of the fact that it does not contain any classical nuclear localization signal (NLS). Using a yeast approach, we observed that the three first LRRs (LRR1-3) and the 32 C-terminal residues of YopM (YopMC-ter) act as NLSs in yeast. Furthermore, by transfection of HEK293T cells, we observed that YopMC-ter could direct large recombinant EGFP-LexA-AD proteins into the nucleus of mammalian cells confirming that it contains a NLS. Critical residues for nuclear targeting were identified by site-directed mutagenesis in YopMC-ter. In addition, we show that YopMC-ter NLS is crucial for the nuclear targeting of an EGFP-YopM fusion protein. PMID- 15043861 TI - Heme-iron increases levels of AP65-mediated adherence by Trichomonas vaginalis. AB - Trichomonas vaginalis is a protozoan responsible for the number one, non-viral sexually transmitted disease. Surface proteins (AP65, AP51, AP33 and AP23) mediate adherence to vaginal epithelial cells (VECs). Iron increases growth of trichomonads and synthesis and surface placement of adhesins. We observed by immunofluorescence using monoclonal antibody (mAb) 12G4 the placement of AP65 on surfaces of trichomonads supplemented with hemoglobin or hemin as a source of iron. We, therefore, tested the hypothesis that heme-bound iron is an alternative source of iron important to trichomonal growth and regulation of expression of the adhesin genes. Here we show that the inhibition of parasite growth by the iron chelator 2,2-dipyridal is rescued by hemoglobin or hemin, but not protoporphyrin IX. Importantly, trichomonads grown in iron-limiting medium supplemented with free iron, hemoglobin and hemin had elevated levels of ap65 transcript that were 12.6-, 12.3- and 9.2-fold higher, respectively, than low iron organisms, as determined by RT-PCR. Similarly, the amounts of AP65 were 8.9 , 11.2-, and 4.8-fold higher in parasites grown in free iron, hemoglobin and hemin, respectively, than organisms in low-iron medium. The heme-iron-regulated AP65 increased adherence of parasites to immortalized VECs. Not surprisingly, parasites pretreated with anti-AP65 serum IgG had decreased adherence compared to organisms incubated with prebleed serum IgG. These data illustrate that heme bound iron is a source of iron similar to lactoferrin. This work extends our findings about the multiple sources of iron for regulating virulence genes of T. vaginalis. PMID- 15043862 TI - Histopathologic changes in kidney and liver correlate with streptococcal pyrogenic exotoxin B production in the mouse model of group A streptococcal infection. AB - Previous studies show that isogenic mutants deficient in streptococcal pyrogenic exotoxin B (SPE B) cause less mortality and skin tissue damage than wild-type strains of Streptococcus pyogenes when inoculated into mice via an air pouch. In this study, the growth and dissemination of bacteria, pathologic changes in various organs, and their correlation with SPE B production were examined. Bacterial numbers in the air pouch from wild-type strain NZ131-infected mice increased at 48 h, while those from speB mutant SW510-infected mice continuously reduced. Mice infected with NZ131 developed bacteremia and greater dissemination in the kidney, liver, and spleen; those infected with SW510 showed either no or slight bacteremia and dissemination. Co-inoculation of SW510 with recombinant SPE B showed a higher bacterial count in the air pouch, bacteremia, and organ dissemination compared to co-inoculation with a C192S mutant lacking protease activity. The histopathologic changes examined showed lesions in kidney and liver in the NZ131-infected but not in SW510-infected mice. The elevation in sera of BUN, AST, and ALT correlated positively with renal and liver impairment. Taken together, SPE B produced during S. pyogenes infection plays a pathogenic role. A direct effect of SPE B on vessel permeability change was also demonstrated. PMID- 15043863 TI - Virulent Burkholderia pseudomallei is more efficient than avirulent Burkholderia thailandensis in invasion of and adherence to cultured human epithelial cells. AB - Burkholderia pseudomallei, a causative agent of melioidosis, is a facultative intracellular gram-negative bacillus that is closely related to its avirulent counterpart, Burkholderia thailandensis. However, pathogenic mechanisms and virulence factors of B. pseudomallei remain elusive. In the present study, we compared the invasiveness, adherence, and replication of B. pseudomallei and B. thailandensis in human respiratory epithelial cells A549. Invasion was determined after 4 h of coculturing using antibiotic protection assay. Adherence was demonstrated by coculturing the cells with fluorescein-labeled bacteria for 1 h and the number of positive cells was analyzed by flow cytometry. The results obtained with this in vitro study demonstrated that compared with its avirulent counterpart, B. pseudomallei is significantly more efficient (P<0.01) in invasion, adherence and inducing cellular damage, as represented by plaque formation. PMID- 15043864 TI - Aromatic amino acid aminotransferase activity and indole-3-acetic acid production by associative nitrogen-fixing bacteria. AB - In this work, we report the detection of aromatic amino acid aminotransferase (AAT) activity from cell-free crude extracts of nine strains of N(2)-fixing bacteria from three genera. Using tyrosine as substrate, AAT activity ranged in specific activity from 0.084 to 0.404 micromol min(-1)mg(-1). When analyzed under non-denaturating PAGE conditions; and using tryptophan, phenylalanine, tyrosine, and histidine as substrates Pseudomonas stutzeri A15 showed three isoforms with molecular mass of 46, 68 and 86 kDa, respectively; Azospirillum strains displayed two isoforms which molecular mass ranged from 44 to 66 kDa and Gluconacetobacter strains revealed one enzyme, which molecular mass was estimated to be much more higher than those of Azospirillum and P. stutzeri strains. After SDS-PAGE, some AAT activity was lost, indicating a differential stability of proteins. All the strains tested produced IAA, especially with tryptophan as precursor. Azospirillum strains produced the highest concentrations of IAA (16.5-38 microg IAA/mg protein), whereas Gluconacetobacter and P. stutzeri strains produced lower concentrations of IAA ranging from 1 to 2.9 microg/mg protein in culture medium supplemented with tryptophan. The IAA production may enable bacteria promote a growth-promoting effect in plants, in addition to their nitrogen fixing ability. PMID- 15043865 TI - Isolation and characterization of an uncoupler-resistant mutant of Methanothermobacter thermautotrophicus. AB - A spontaneous mutant of Methanothermobacter thermautotrophicus resistant to the protonophorous uncoupler TCS was isolated. The mutant strain exhibited increased CH(4) formation and elevated level of ATPase activity under non-growing conditions. ATP synthesis driven by methanogenic electron transport as well as by potassium diffusion potential in the presence of either H(+) or Na(+) ions was markedly diminished in the mutant strain. An abundant membrane-associated protein complex with molecular mass approximately 670 kDa was detected in the mutant strain after native PAGE. The results indicate that TCS resistance in this mutant has arisen as a consequence of mutation(s) that affects a specific locus coding for an uncoupler binding protein(s) and/or modulate the activity of unidentified ATPase. PMID- 15043866 TI - Involvement of caspase activation through release of cytochrome c from mitochondria in apoptotic cell death of macrophages infected with Actinobacillus actinomycetemcomitans. AB - We previously reported that infection with the periodontopathic bacterium Actinobacillus actinomycetemcomitans induced apoptosis in a mouse macrophage cell line J774.1. In the present study, we examined the involvement of cytochrome c and caspases in the induction of apoptosis in A. actinomycetemcomitans-infected J774.1 cells. Following infection, the expression levels of cytochrome c, and cleaved forms of caspase-3 and caspase-9 in the cells were examined using immunoblot analysis. Cytochrome c was released from mitochondria into the cytoplasm after A. actinomycetemcomitans-infected J774.1 cells were cultured for 6 h, and caspase-3 and caspase-9 were found to be cleaved forms in the cells. Further, caspase-9 activity was markedly increased, and phosphorylated p53 was detected in the cells 30 h following infection. These results suggest that apoptosis in A. actinomycetemcomitans-infected J774.1 cells is regulated by the release of cytochrome c from mitochondria into cytoplasm and the subsequent activation of caspases through phosphorylation of p53. PMID- 15043867 TI - Distribution and composition of the lysis cassette of Lactococcus lactis phages and functional analysis of bacteriophage ul36 holin. AB - The bacteriophage lysis cassette, which comprises a lysin and a holin gene, was analyzed in 18 Lactococcus lactis phages. A muramidase motif was found in the lysins of c2-like phages, while an amidase motif was observed in the lysins of 936-like phages. Both amidase and muramidase types were detected among the P335 phages. The P335 lysins were separated into three groups based on amino acid sequence identity. A class I holin was recognized in 936-like and c2-like phages, whereas P335-like phages possess class II holins. The P335 holins were further divided into four groups based on sequence identity. Only the holins of 936-like phages contained putative dual-start motifs. The unusual lysis cassette of the highly virulent P335-like phage ul36 contains a unique holin (orf74B) upstream of a lysin which is present in several other P335-like phages. Using the lambdadelta Sthf system, we demonstrated that gpORF74B induces cell lysis at the same time as lambdadelta Sthf::S105, the effector of lambda lysis. Transcriptional analysis of ul36 lysis cassette showed that first transcripts are detected 35 min after infection of L. lactis cells. The lysis clock of phage ul36 appears to be controlled by the late expression of the holin and lysin genes. PMID- 15043868 TI - Distribution of virulence genes of Staphylococcus aureus isolated from stable nasal carriers. AB - In the present study, we report data on virulence determinants of Staphylococcus aureus from stable nasal carriers, emphasizing on the genes encoding fibronectin (fnbA, fnbB) and collagen (cna) adhesive molecules. Of the 44 S. aureus isolates included, 32 isolates (16 pairs) were cultured from the anterior nares of 16 healthy carriers, eight isolates (four pairs) were collected from the nose of four patients with recurrent skin infections and four isolates were obtained from the infection site of these patients. The period between the two nasal swabs taken was 3-5 days. The persistency of carriage could be demonstrated by the indistinguishable genotypic characteristics of the S. aureus isolates in each pair. This could be shown by determination of gene polymorphisms of coa gene and the X-region and IgG-binding region encoding segments of spa gene. In addition, the isolates within the pairs showed identical toxin patterns. This was determined by PCR amplification of the genes encoding staphylococcal enterotoxins (SEA to SEJ) and TSST-1. The genotypic properties also yielded an identity between persistent nasal carriage isolates and the corresponding skin infection isolates of the four patients. In addition, all S. aureus nasal and skin infection isolates were positive for gene fnbA, fnbB and cna could be found with a high frequency. Among the 44 isolates investigated, 16 isolates (36.7%) harbored gene fnbB and 21 isolates (47.7%) gene cna. The data in the present study showed a relatively wide distribution of the genes fnb and cna among the investigated isolates, indicating that the persistent carriage of strains harboring these virulence determinants may increase the risk for subsequent invasive infections in carriers. PMID- 15043869 TI - Cloning and expression of Lipomyces starkeyi alpha-amylase in Escherichia coli and determination of some of its properties. AB - The Lipomyces starkeyi alpha-amylase (LSA) gene encoding soluble starch-degrading alpha-amylase was cloned and characterized from a derepressed and partially constitutive mutant for both dextranase and amylase activities. The nucleotide (nt) sequence of the cDNA fragment reveals an open reading frame of 1944 bp encoding a 619 amino acid (aa) mature protein (LSA) with a calculated molecular weight of 68.709 kDa that was estimated to be about 73 kDa, including His tag (4 kDa) based on SDS-PAGE (10% acrylamide gel), activity staining, and the Western blotting, using anti-amylase-Ab. LSA had a sequence similar to other alpha amylases in four conserved regions of the alpha-amylase family: (I) (287)DIVVNH(292), (II) (372)GLRIDTVKH(380), (III) (399)GEVFD(403), (IV) (462)FLENQD(467). Polymerase chain reaction and sequence analysis showed one intron of 60 nucleotides in the genomic lsa at positions between 966 and 967 of cDNA. The cloned LSA amylase showed a maximum activity at pH 6 and optimum temperature of 40 (o)C, with greater than 90% stability between pH 5 and pH 8 for 16 h. It was inhibited by Cu(2+) and stimulated by Ca(2+) and Mg(2+). Enzyme activity was not affected by 1 mM EGTA but was inhibited by 1 mM EDTA. LSA did not hydrolyze maltodextrins of G2 to G4, yet formed G2+G3 from G5, G2+G4 or G3+G3 from G6, and G3+G4 from G7. LSA did not hydrolyze soluble starch in the present of 2% (w/v) of acarbose. Kinetics of LSA was carried out by using starch as a substrate and the inhibition type of acarbose was the mixed non-competitive type (ki = 3.4 microM). PMID- 15043870 TI - Phospholipids of Clostridium perfringens: a reexamination. AB - We have identified phosphatidylethanolamine as one of the major phospholipids of Clostridium perfringens by two dimensional thin layer chromatography of the intact lipids and of their deacylation products and by liquid chromatography followed by mass spectrometry of the intact neutral phospholipid fraction. The principal fatty acids of phosphatidylethanolamine are myristic acid (14:0), lauric acid (12:0), and palmitic acid (16:0) and the major molecular species are 14:0,14:0 (26.3%); 12:0,14:0 (19.0%); 14:0,16:0 (22.4%) and 16:0,16:0 (17.6%). A similar distribution of molecular species was found in the other major phospholipid, O-alanyl phosphatidylglycerol. PMID- 15043871 TI - Development of PCR assays for the detection and differentiation of Fusarium sporotrichioides and Fusarium langsethiae. AB - Isolates of the type-A trichothecene producing Fusarium sporotrichioides and Fusarium langsethiae were grouped and differentiated in a phylogenetic tree using ITS sequence dissimilarity. An attempt was made to develop a PCR-based assay for the detection and differentiation of Fusarium sporotrichiodes from other Fusarium species using the 5'-region of the tri5 gene as a template. However, this assay was unable to differentiate, to a satisfactory level, between isolates of Fusarium sporotrichioides and Fusarium langsethiae, providing further genetic evidence for their close genetic relationship. A robust and repeatable PCR-assay was developed for the detection and differentiation of both species based on sequence determined from differentially amplified RAPD-PCR products. These assays were able to detect both species in samples of grain taken from the field. PMID- 15043872 TI - Analysis of microbial diversity in oligotrophic microbial fuel cells using 16S rDNA sequences. AB - Molecular ecological techniques were applied to analyze the bacterial diversity of two oligotrophic microbial fuel cells (MFCs) enriched using river water or artificial wastewater (AWW) as fuel. Denaturing gradient gel electrophoresis (DGGE) of the PCR amplified 16S rDNA showed that different microbial communities were present in the two MFCs and these were different from the river sediment used to initiate the enrichment. Nearly complete 16S rDNA was amplified and sequenced. Over 80% of the clones were Proteobacteria. Betaproteobacteria were the dominant clones (46.2%) in MFCs fed with river water, and about 64.4% of the clones in MFCs fed with AWW were Alphaproteobacteria. Actinobacteria were found only in the MFC fed with AWW, and Deltaproteobacteria, Acidobacteria, Chloroflexi and Verrucomicrobia in the MFC fed with river water. Many clones were related to uncultured bacteria, some with homology less than 95%, indicating that many novel bacteria were enriched in the oligotrophic MFCs. PMID- 15043873 TI - Identification of the anginosus group within the genus Streptococcus using polymerase chain reaction. AB - The aim of this study was to establish an identification method for the anginosus group within the genus Streptococcus by polymerase chain reaction (PCR). Using a primer pair based on the group-specific sequences of penicillin-binding protein 2B (pbp2b) gene, a 275-bp fragment was amplified from each species in the group but no size-matched products were obtained in other streptococci. Further identification in the species or subspecies level was possible by a multiplex PCR with primers for the 16S ribosomal RNA gene of Streptococcus anginosus, the hyaluronate lyase genes both of Streptococcus intermedius and Streptococcus constellatus subsp. constellatus, and the intermedilysin (ily) gene of S. intermedius. In the case ofStreptococcus constellatus subsp. pharyngis, the amplified fragment from the S. intermedius-type hyaluronate lyase gene was obtained, while that from the ily gene was not. These results also indicate that two different hyaluronate lyase genes are distributed among the anginosus group. PMID- 15043875 TI - Genome-wide cloning and characterization of microbial esterases. AB - We have isolated putative esterase genes from various bacterial chromosomes. Thirty open reading frames predicted to encode esterases were randomly selected from 13 sequenced bacterial chromosomes and were cloned into an expression vector. The esterase activity of the resulting clones was tested on a tributyrin plate at different pH values and temperatures. Nine out of thirty tested clones exhibited significant tributyrin hydrolyzing activity. The enzyme S5 from the gene b0494 of Escherichia coli, the enzyme S12 from the gene STM0506 of Salmonella typhimurium, and the enzyme S28 from the gene AF1716 of Archaeoglobus fulgidus exhibited high activity at an alkaline pH range. The esterase S11 encoded by the gene PA3859 of Pseudomonas aeruginosa PAO1 and the esterase S21 from the gene SMc01033 of Sinorhizobium meliloti 1021, both showed a sharp increase in enzyme activity above pH 8.0. Furthermore, the enzymes S5, S12, S21, and S28 retained the esterase activity when they were incubated at 50 degrees C, suggesting that these enzymes are thermostable. Subsequent pH vs. activity and temperature vs. activity experiments with selected enzymes in a solution assay system confirmed the validity of the above data. The genome-wide exploration strategy of proteins provided valuable information on the esterases by revealing subtle biochemical differences between the esterases of different sources. PMID- 15043874 TI - Centrin in Giardia lamblia - ultrastructural localization. AB - Giardia lamblia is a multiflagellar parasite and one of the earliest diverging eukaryotic cells. It possesses a complex cytoskeleton based on different groups of microtubular structures - a ventral adhesive disc, four pairs of flagella, a median body and funis. Centrin is an important member of the EF-hand family of calcium-binding proteins, and it is known to show calcium-sensitive contractile behaviour. In the present study, we performed an ultrastructural localization of centrin in G. lamblia using several monoclonal antibodies to centrin. Microtubular structures such as the basal bodies, all the flagella axonemes, the adhesive disc, funis, and the median bodies presented positive labelling to centrin. In addition, the dense rods also demonstrated positive labelling. These results show that centrin is located in key positions related to microtubules. The role of centrin in these dynamic regions is discussed. PMID- 15043877 TI - Similarity of bacterial communities in sawdust- and straw-amended cow manure composts. AB - We analyzed bacterial communities in two cow manure composts derived from the same feed manure and composted in the same location, but composted with different carbon amendments, and in peat-based potting mixes amended with these composts. Bacterial communities were characterized by PCR-denaturing gradient gel electrophoresis (DGGE) analysis of extracted DNAs, and population fingerprints generated for each sample were compared. Sequence analyses of dominant DGGE bands revealed that members of the phylum Bacteroidetes were the most dominant bacteria detected in this study (19 of 31 clones). These analyses demonstrate that bacterial community profiles of individual composts were highly similar, as were profiles of compost-amended potting mixes. However, potting mix profiles differed substantially from the original compost profiles and from that of the peat base. These data indicate that highly similar bacterial populations were active in the two composts, and suggest that the effects of the initial carbon amendment on the mature compost bacterial communities were minor, while factors such as the feed manure and composting location may have been more influential. PMID- 15043878 TI - Oxygen flux magnitude and location along growing hyphae of Neurospora crassa. AB - Oxygen fluxes were mapped at the growing apices and along mycelial hyphal segments of the ascomycete Neurospora crassa. High spatial resolution was obtained using micro-oxygen probes (2-3 microm tip diameters) and the self referencing technique to maximize the sensitivity of oxygen flux measurements. As expected, oxygen influx was inhibited by cyanide, although oxygen influx (and hyphal growth) resumed with the induction of an alternate oxidase activity. Along hyphal segments, variations in oxygen influx were not correlated with location, near or far from septa, and varied over time along the same hyphal segment. Growing hyphae had a region of maximal oxygen influx greater than 10 microm behind the hyphal tip, the oxygen influx was correlated with hyphal growth rate. The region of maximal oxygen influx did not correspond with mitochondrial density, which is maximal (about 30% of hyphal volume) 5-10 microm behind the tip. Therefore, tip-localized mitochondria do not contribute to the respiratory requirements of the growing hypha. The tip-localized mitochondria may function in clearing calcium from the cytoplasm, although a decline in chlortetracycline fluorescence after cyanide inhibition could also be due to ATP-depletion due to inhibition of actively respiring mitochondria. Alternatively, the growing tip may be the site of mitochondrial biogenesis. PMID- 15043876 TI - PknH, a transmembrane Hank's type serine/threonine kinase from Mycobacterium tuberculosis is differentially expressed under stress conditions. AB - Serine/threonine protein kinases (STPKs) represent a burgeoning concept in prokaryotic signaling and have been implicated in a range of control mechanisms. This paper describes the enzymatic and molecular characterization of PknH, a mycobacterial STPK. After cloning and expression as a Glutathione-S-transferase fusion protein in E. coli, PknH was found to phosphorylate itself and exogenous substrates like myelin basic protein and histone. The kinase activity of PknH was inhibited by the kinase inhibitors staurosporine and H-7. The results confirmed that PknH is a transmembrane protein and is restricted to members of the Mycobacterium tuberculosis complex. In addition, transcriptional analysis of pknH in M. tuberculosis under various stress conditions revealed that exposure to low pH and heat shock decreased the level of pknH transcription significantly. This is the first report describing differential expression of a mycobacterial kinase in response to stress conditions which can indicate its ability to regulate cellular events promoting bacterial adaptation to environmental change. PMID- 15043879 TI - Knockout of the alanine racemase gene in Lactobacillus plantarum results in septation defects and cell wall perforation. AB - A stable mutant of Lactobacillus plantarum deficient in alanine racemase (Alr) was constructed by two successive homologous recombination steps. When the mutant was supplemented with D-alanine, growth and viability were unaffected. Surprisingly, deprivation of d-alanine during exponential growth did not result in a rapid and extensive lysis as observed in Alr-deficient strains of Escherichia coli or Bacillus subtilis. Rather, the starved mutant cells underwent a growth arrest and were gradually affected in viability with a decrease in colony forming units over 99% in less than 24 h. Additionally, fluorescent techniques demonstrated a loss of cell envelope integrity in the starved cells. Prolonged d-alanine starvation resulted in cells with an aberrant morphology. Scanning and transmission electron microscopy analyses revealed an increase in cell length, deficiencies in septum formation, thinning of the cell envelope and perforation of the cell wall in the septum region. We discuss the involvement of peptidoglycan hydrolases in these phenotypic defects in the context of the crucial role played by D-alanine in peptidoglycan biosynthesis and teichoic acids substitution. PMID- 15043881 TI - Virulence of Streptococcus pneumoniae may be determined independently of capsular polysaccharide. AB - Mice were inoculated intranasally with Streptococcus pneumoniae isolates of serotype 14 with different genetic backgrounds (14R, 14DW) and a capsular switch of 14R, strain 9VR (serotype 9V). Inoculation of the mice with 14R and 9VR resulted in 60% mortality. All the mice survived 14DW inoculation. No differences in lungs' bacterial loads were found 3 h following inoculation. Bacterial clearance of 5 logs was observed 48 h after inoculation with 14DW versus within 1 log 48 h after inoculation with 14R and 9VR. No significant differences in bacterial size or the capsular amount could be found between 14R and 14DW. We conclude that factor(s) in addition to the capsule, contribute to disease outcome. PMID- 15043880 TI - Induction of baroresistance by hydrogen peroxide, ethanol and cold-shock in Saccharomyces cerevisiae. AB - The acquisition of tolerance to high hydrostatic pressure of 220 MPa (HHP) in response to a 0.4 mM hydrogen peroxide, 6% ethanol and cold-shock (10 degrees C) pretreatment for different lengths of times was studied in the yeast Saccharomyces cerevisiae. The protection conferred by these different treatments was similar ( approximately 3 log cycles) and time-dependent. Analysis of the induction of the most pressure up-regulated genes under these conditions was investigated by RT-PCR. Our results revealed that the cell stress response to HHP shares common features with hydrogen peroxide and ethanol stresses, but differs in some way to cold-shock. PMID- 15043883 TI - Listeria monocytogenes: comparative interpretation of mouse virulence assay. AB - Being an opportunistic bacterial pathogen, Listeria monocytogenes demonstrates significant strain variations in virulence and pathogenicity. The availability of laboratory procedures to ascertain the pathogenic potential of L. monocytogenes bacteria would greatly enhance the control and prevention of listerial infections. As a method that measures all virulent determinants, mouse virulence assay has been frequently used for assessing L. monocytogenes virulence. The pathogenic potential of a given L. monocytogenes strain as determined by mouse virulence assay is often calculated from mouse mortality data in combination with colony forming units (CFUs) derived from plate counts, and expressed by medium lethal dose (LD(50)). In this report, we describe an alternative method [i.e., relative virulence (%)] that does not involve CFU estimation, and is comparable to LD(50) for interpretation of mouse virulence assay for L. monocytogenes. The relative virulence (%) is obtained by dividing the number of dead mice with the total number of mice tested for a particular strain using a known virulent strain (e.g., L. monocytogenes EGD) as reference. Besides providing a more direct interpretation in comparison with LD(50) values for mouse virulence assay, this method requires fewer dosage groups per L. monocytogenes strain, and eliminates CFU estimation that is step subject to variations between runs and also between laboratories. PMID- 15043882 TI - The non-photosynthetic, pathogenic green alga Helicosporidium sp. has retained a modified, functional plastid genome. AB - A fragment of the Helicosporidium sp. (Chlorophyta: Trebouxiophyceae) plastid genome has been sequenced. The genome architecture was compared to that of both a non-photosynthetic relative (Prototheca wickerhamii) and a photosynthetic relative (Chlorella vulgaris). Comparative genomic analysis indicated that Helicosporidium and Prototheca are closely related genera. The analyses also revealed that the Helicosporidium sp. plastid genome has been rearranged. In particular, two ribosomal protein-encoding genes (rpl19 and rps23) appeared to have been transposed, or lost from the Helicosporidium sp. plastid genome. RT-PCR reactions demonstrated that the retained plastid genes were transcribed, suggesting that, despite rearrangement(s), the Helicosporidium sp. plastid genome has remained functional. The modified plastid genome architecture is a novel apomorphy that indicates that the Helicosporidia are highly derived green algae, more so than Prototheca spp. As such, they represent a promising model to study organellar genome reorganizations in parasitic protists. PMID- 15043884 TI - Differences in malate dehydrogenases from the obligately piezophilic deep-sea bacterium Moritella sp. strain 2D2 and the psychrophilic bacterium Moritella sp. strain 5710. AB - The gene encoding malate dehydrogenase (MDH) of the obligately piezophilic deep sea bacterium Moritella sp. strain 2D2 was cloned and sequenced. There were two positions [close to the active site (Ala-180) and in the subunit interaction site (His-229)] with 2D2-specific substitutions. The MDH genes of strain 2D2 and a psychrophilic bacterium Moritella sp. strain 5710 exhibiting the highest sequence similarity were overexpressed in Escherichia coli. The 2D2 MDH was more heat stable than the 5710 MDH. The apparent Km value at 62.1 MPa for NADH of the 2D2 MDH was higher than that of the 5710 MDH. The 2D2 MDH in which a His-Gln substitution was introduced at position 229 decreased the thermal stability and Km value at 62.1 MPa. The 5710 MDH that was substituted Gln-229 with His increased the thermal stability and Km value at 62.1 MPa. These results indicate that the His residue at position 229 of the 2D2 MDH may play a role in the thermal stability and the MDH function at high pressure. PMID- 15043886 TI - Silas Weir Mitchell, MD: the physician who discovered causalgia. AB - Silas Weir Mitchell discovered and treated causalgia, a condition most often encountered by hand surgeons. He is considered the father of neurology as well as an early pioneer in scientific medicine. He was also a psychiatrist, toxicologist, author, poet, and a celebrity in America and Europe. His many skills and interests led his contemporaries to consider him a genius on par with Benjamin Franklin. His contributions to medicine and particularly hand surgery continue to resonate today. PMID- 15043885 TI - Characterisation of Shiga toxin-producing Escherichia coli (STEC) isolated from seafood and beef. AB - Shiga toxin-producing Escherichia coli (STEC) strains isolated in Mangalore, India, were characterised by bead-enzyme-linked immunosorbent assay (bead-ELISA), Vero cell cytotoxicity assay, PCR and colony hybridisation for the detection of stx1 and stx2 genes. Four strains from seafood, six from beef and one from a clinical case of bloody diarrhoea were positive for Shiga toxins Stx1 and Stx2 and also for stx1and stx2 genes. The seafood isolates produced either Stx2 alone or both Stx1 and Stx2, while the beef isolates produced Stx1 alone. The stx1 gene of all the beef STEC was found to be of recently reported stx1c type. All STEC strains and one non-STEC strain isolated from clam harboured EHEC-hlyA. Interestingly, though all STEC strains were negative for eae gene, two STEC strains isolated from seafood and one from a patient with bloody diarrhoea possessed STEC autoagglutinating adhesion (saa) gene, recently identified as a gene encoding a novel autoagglutinating adhesion. PMID- 15043887 TI - Giant-cell tumor of the distal forearm. AB - PURPOSE: Many authorities express concern that giant-cell tumors of the distal forearm are more frequently recurrent and difficult to treat chiefly because of the proximity to the carpus and the resultant diminished range of motion in the hand and forearm. We have studied the results from our institution for 49 patients with giant-cell tumors of the distal forearm treated from 2 to 28 years (mean, 14 +/- 7 years) and compared the results for different methods of treatment. METHODS: Through the computer database 49 patients with giant-cell tumors of the distal forearm (46 of the radius, 3 of the ulna) were identified. By using material from patient visits, chart review, and when necessary telephone interviews it was possible to gather demographic and outcome data for the 49 patients. For the radial lesions, 15 of the patients had a marginal resection of the distal radius along with the periosteum and ligamentous structures and implantation of cadaveric allografts. Twenty-six patients had intralesional curettage and insertion of polymethylmethacrylate (PMMA) and 5 had curettage and autograft insertion. The 3 patients with ulnar lesions were treated with Darrach resections. RESULTS: There were no deaths, infections, metastases, or amputations. In addition to the 49 original surgeries, the patients required 41 additional surgical procedures, 17 of which were for recurrent disease. The greatest numbers of recurrences were in the patients who underwent curettage with autograft or PMMA implantation. Only 2 recurrences were in patients who had a marginal resection and implantation of cadaveric allografts. The overall results for the patients showed that many had mostly minor complaints referable to function or pain and that only 18 of the 49 patients were asymptomatic. CONCLUSIONS: Although the patients with distal forearm giant-cell tumors have had a difficult course in terms of local recurrence and subsequent surgeries required for treatment failures, the ultimate outcomes for both allograft transplantation and curettage and insertion of PMMA are satisfactory. The patients with complete distal radial allografts had a better record for prevention of recurrence than the patients treated with curettage and PMMA insertion but the percentages of currently asymptomatic patients are approximately the same for both series (40% [6/15], 35% [9/26]). These data support the concept that marginal resection and complete distal radial allograft implantation should be used for patients with tumors that have destroyed much of the bone and have extensive soft tissue components and that curettage and PMMA insertion should be reserved for patients where the structural alteration of the bone is minimal. PMID- 15043888 TI - Malignant fibrous histiocytoma of the distal radius: a case report. AB - We report a case of a primary malignant fibrous histiocytoma of the distal radius in a 46-year-old man. Treatment involved en bloc resection, reconstruction with a nonvascularized free fibular autograft, and wrist arthrodesis combined with adjuvant chemotherapy. At the 2-year follow-up evaluation the patient had a satisfactory outcome with complete radiographic union and no evidence of a local recurrence or metastasis. Resection combined with autogenous fibular grafting and adjuvant chemotherapy appears to be an acceptable method for treating malignant fibrous histiocytoma of the distal radius in this patient. PMID- 15043889 TI - Keloid formation after syndactyly reconstruction: associated conditions, prevalence, and preliminary report of a treatment method. AB - PURPOSE: The purpose of this study is 3-fold: to review our cases of keloid formation after syndactyly release, to report a clinical association between primary enlargement of the digits and risk of keloid formation, and to report treatment using low-dose, short-term methotrexate as an adjunct to revision surgery. METHODS: A retrospective review of patients identified with keloid formation after syndactyly reconstruction showed associated enlargement of the involved digits. A search of medical records for cases in which both syndactyly and digital enlargement occurred was carried out. Charts and images, where available, were reviewed for information about age, gender, involved site, associated conditions, and treatment. RESULTS: Eight cases of keloids occurred in 1004 surgical procedures in 681 patients carried out during the 20-year period reviewed. Seven patients were white and 1 was Hispanic. Seven of the 8 had associated primary digital enlargement. Two patients who had enlarged syndactylized digits did not develop keloids after surgery. There was no family history in any case. Standard treatment (pressure, topical or intralesional corticosteroids, and re-excision) was unsuccessful in resolving the keloids. Two children treated with adjunctive methotrexate had successful treatment of their keloids and near-normal healing. Length of follow-up time after the last treatment ranged from 6 months to 11 years (average, 5.5 years). CONCLUSIONS: Primary digital enlargement is highly predictive of risk of keloid formation after syndactyly reconstruction. Very-low-dose, short-term methotrexate was successful as an adjunct to surgical treatment in 2 cases. PMID- 15043890 TI - The abdominal flap using scarred skin in the treatment of postburn hand deformities of severe burn patients. AB - PURPOSE: To present a random-pattern flap technique using scarred skin from the abdominal wall to cover the defect after burn scar was released from the hand and the scarred skin raised from the hand was transferred to the abdominal wall to cover the flap-donor area in patients who suffered from severe burns involving the entire body and had no available healthy skin to be used for coverage. METHODS: Ten hands of 7 patients (5 males, 2 females) were treated in our department between April 1994 and February 2001. The total body surface area involved with second-or third-degree burns was 85% to 96% and there was no available healthy skin to be used for reconstruction. All hands had severe scar contracture on the dorsum and lost most of their function and the patients lost the ability to take care of themselves. The scarred skin on the dorsum of the hand was elevated integrally as a flap based on the ulnar border of the hand. The tendons were released, the stiff metacarpophalangeal joints were mobilized by closed capsulotomies, and the contracture of the thumb web space was released. A random-pattern abdominal flap using scarred skin was designed and elevated to cover the defect of the scar-released hand and the scarred skin raised from the dorsum of the hand was transferred to cover the defect of the abdominal flap donor site. RESULTS: The flaps that were transferred to the dorsum of the hands survived entirely and the defects of the flap donor area were covered perfectly without further skin graft. All patients were evaluated for 0.5 to 4 years after surgery. The range of motion of the metacarpophalangeal joints and the space capacity of the thumb web were improved greatly. All patients regained the ability to take care of themselves. CONCLUSIONS: This method is simple and has satisfactory results. It appears from our experience that the mature postburn scarred skin on the abdomen can be used for a flap procedure for reconstructing the burned hand and that the scarred skin raised from the dorsum of the hand can be transferred to cover the flap donor sites on the abdomen. Under circumstances in which there is no normal skin for reconstruction the function of the burned hand could be improved greatly by this method. PMID- 15043891 TI - Central slip tenotomy for the treatment of chronic mallet finger: an anatomic study. AB - PURPOSE: Tenotomy of the central slip, described by Fowler, can clinically improve chronic distal interphalangeal joint (DIP) extensor lag secondary to mallet finger (terminal tendon disruption). The goal of this study is to evaluate the potential of central slip tenotomy to restore DIP joint extension. METHODS: A mallet deformity was reproduced in 15 fresh-frozen cadaver fingers after the extensor tendon insertion was sectioned over the DIP joint. A suture anchor inserted at the terminal insertion was then secured to the extensor tendon over the middle phalanx to reconstruct the extensor mechanism. A 500-g weight attached to the proximal extensor tendon applied extensor tension. Central slip tenotomy was then performed. DIP extensor lags before and after tenotomy were recorded. RESULTS: After sectioning of the terminal tendon over the DIP joint the average amount of extensor tendon lag produced was 45 degrees. After central slip tenotomy was performed the average amount of extensor lag correction was 36 degrees (range, 30 degrees-46 degrees). CONCLUSIONS: Several clinical studies have shown that central slip tenotomy is an effective treatment for chronic mallet finger but may not fully restore DIP joint extension. Our data suggest that patients with a pre-existing extensor lag of greater than 36 degrees may not achieve full extension from central slip tenotomy, although extensor lags of up to 46 degrees may be corrected. PMID- 15043892 TI - Measurement of gliding resistance of the extensor pollicis longus and extensor digitorum communis II tendons within the extensor retinaculum. AB - PURPOSE: The etiology of spontaneous extensor pollicis longus (EPL) tendon rupture is still largely unknown. It is possible that friction within the sheath may play a role. The purposes of this study were to compare gliding resistance of the EPL tendon with that of the extensor digitorum communis tendon of the index finger (EDC II) and to find the wrist position that gives the EPL tendon the lowest gliding resistance. METHODS: Fifteen fresh-frozen cadavers were used. Gliding resistance was measured directly in 7 different wrist positions. RESULTS: The mean gliding resistance of the EPL tendon was 0.16 +/- 0.08 N and that of the EDC II tendon was 0.11 +/- 0.06 N. This difference was significant. There was also a significant effect on gliding resistance due to wrist position. For the EPL tendon, the gliding resistance was significantly greater in 60 degrees wrist flexion compared with all other wrist positions tested. Additionally the gliding resistance of the EPL in 30 degrees flexion, 60 degrees extension, and 15 degrees radial deviation was significantly higher than wrist positions of 30 degrees extension, neutral, and 30 degrees ulnar deviation. CONCLUSIONS: Positioning the wrist close to neutral flexion/extension and in some ulnar deviation minimizes the friction within the EPL sheath. Such positions may be advantageous for splinting patients at risk for EPL rupture. PMID- 15043893 TI - Extensor pollicis longus tenosynovitis mimicking de Quervain's disease because of its course through the first extensor compartment: a report of 2 cases. AB - We present 2 cases of extensor pollicis longus tenosynovitis caused by the unusual course of the extensor pollicis longus tendon. The extensor pollicis longus tendon passed through the first extensor compartment and was constricted severely. The symptoms were similar to those of de Quervain's disease. The clinical presentation, diagnosis, and surgical technique of this very rare anatomic condition are discussed. PMID- 15043894 TI - Inhibition of TGF-beta-induced collagen production in rabbit flexor tendons. AB - PURPOSE: Postoperative adhesions frequently compromise the success of flexor tendon repair. Manipulation of growth factors responsible for scar formation may be a method of decreasing adhesion formation. Transforming growth factor beta (TGF-beta) is a key cytokine in the pathogenesis of tissue fibrosis. The purpose of this study was to examine the effectiveness of TGF-beta neutralizing antibody in blocking TGF-beta-induced collagen I production in rabbit flexor tendons in vitro. METHODS: Sheath fibroblasts, epitenon tenocytes, and endotenon tenocytes were obtained from rabbit flexor tendons. Each cell culture was supplemented with 1 ng/mL of TGF-beta along with increasing doses of TGF-beta neutralizing antibody (0.1-2.0 microg/mL). Collagen I production was measured by enzyme-linked immunoabsorbent assay and TGF-beta bioactivity was measured by the luciferase assay. Results were compared with TGF-beta alone and unsupplemented controls. RESULTS: The addition of neutralizing antibody significantly reduced TGF-beta induced collagen I production in a dose-dependent manner in all 3 cell cultures. TGF-beta bioactivity was also reduced by its neutralizing antibody. CONCLUSIONS: This study shows that TGF-beta inhibition through its neutralizing antibody was effective in cultured flexor tendon cells. The results encourage further experiments that use such agents to modulate flexor tendon wound healing in in vivo models in the hope of eventually blocking the effect of TGF-beta on flexor tendons clinically. PMID- 15043895 TI - Pisotriquetral instability causing an unusual flexor tendon rupture. AB - We present a case of a closed rupture of the flexor digitorum profundus tendon of the small finger. It is our hypothesis that because of pisotriquetral instability as a result of a lesion of the pisotriquetral ligaments the flexor digitorum profundus tendon was entrapped repeatedly between the pisiform and triquetrum. This resulted in an attritional lesion of the tendon. The pisotriquetral joint showed no bone abnormalities. PMID- 15043896 TI - Ligament reconstruction with trapezium retention arthroplasty for carpometacarpal arthritis. AB - PURPOSE: The purpose of this study is to provide a qualitative and quantitative assessment of results in a consecutive series of patients with carpometacarpal arthritis using a trapezium-retaining interposition arthroplasty. METHODS: Between 1999 and 2002, 16 patients with basal joint arthritis were treated with a trapezium-retaining interposition arthroplasty. Fourteen of these patients were evaluated at a mean follow-up time of 20 months (minimum, 12 months) and were compared based on clinical outcome, presurgical and postsurgical x-ray evaluation, and range of motion, grip-strength, and pinch-strength values. RESULTS: Average trapezial height values as quantified by the trapezial space to proximal phalanx ratio were very similar before and after surgery. Average grip strength values improved significantly. Although average pinch-strength and average range of motion values improved neither of these comparison values were significant. Thirteen of the 14 patients (93%) were satisfied with their results as quantified by postsurgical Disabilities of the Arm, Shoulder, and Hand questionnaires. CONCLUSIONS: Ligament reconstruction with trapezium retention arthroplasty of the basal joint not only successfully maintains trapezial height but also yields patient satisfaction rates comparable to those achieved with other forms of basal joint arthroplasty. PMID- 15043897 TI - Basal joint osteoarthritis of the thumb: a prospective trial of steroid injection and splinting. AB - PURPOSE: There have been few prospective studies evaluating the results of nonsurgical treatment of a well-defined patient cohort with symptomatic basal joint osteoarthritis of the thumb. This prospective study uses a validated outcome instrument to examine the effectiveness of a single steroid injection and 3 weeks of splinting in patients with osteoarthritis in Eaton stages 1 to 4 with a minimum of 18 months of follow-up evaluation. METHODS: Thirty consecutive patients (30 thumbs) were studied prospectively to evaluate the efficacy of a single injection of corticosteroid into the trapeziometacarpal joint, followed by immobilization in a thumb spica splint for 3 weeks. All patients answered an outcome-based questionnaire (Disabilities of the Arm, Shoulder, and Hand) and were examined before injection, 6 weeks after injection, and at final follow-up examination (minimum, 18 months). Eaton radiographic stage was recorded by 3 independent observers. RESULTS: At 6 weeks 13 patients had improvement in pain intensity and 17 patients reported no symptomatic improvement. Twelve of those with relief at 6 weeks continued to have relief at long term follow-up evaluation (mean, 25 months). Of patients with long-term relief average grip strength of the affected thumb was 95% of contralateral side, whereas those without relief had grip strength values that were 60% of contralateral side. For those patients without relief at 6 weeks there was no improvement seen at later follow-up evaluation. Five patients with Eaton stage 1 disease had an average of 23 months of relief with nonsurgical treatment. In stage 2 and stage 3 disease 7 thumbs improved at 6 weeks after injection and 6 thumbs had long-term relief. In stage 4 disease, 6 thumbs had neither short-term nor long-term relief with the injection. Disease side, handedness, and smoking did not affect outcomes. At final follow-up evaluation 12 thumbs had had surgical treatment. CONCLUSIONS: Steroid injection with splinting for the treatment of basal joint arthritis of the thumb provided reliable long-term relief in thumbs with Eaton stage 1 disease but provided long term relief in only 7 of 17 thumbs with Eaton stage 2 and stage 3 basal joint arthritis. PMID- 15043898 TI - Comparative in vivo kinematic analysis of normal and osteoarthritic trapeziometacarpal joints. AB - PURPOSE: Although extensive studies have been conducted on the trapeziometacarpal (TM) joint, there have been few quantitative studies on its in vivo kinematics. The purposes of this study are to quantitatively evaluate the in vivo kinematics of the TM joint with a fluoroscope and to determine the difference between normal and arthritic joints. METHODS: Fourteen healthy hands and 8 symptomatic hands with idiopathic osteoarthritis were included in this study. The healthy subjects were divided into 2 groups: a younger control group aged under 50 years (n = 7) and an older control group aged 51 years or older (n = 7). Radial and dorsal translations of the center of rotation (COR) were quantified in active abduction adduction and active flexion-extension. In addition, volar tilt of the metacarpal facet of the TM joint was measured on the lateral view. RESULTS: The CORs in flexion-extension translated more dorsally in the patient group than those in the 2 control groups whereas the CORs in abduction-adduction did not differ significantly between the patient group and the control groups. There was no significant difference in the translation of COR between the 2 control groups in either abduction-adduction or flexion-extension. The average volar tilt in the patients with osteoarthritis increased significantly compared with those in the control groups. CONCLUSIONS: Dorsal translation of the COR in flexion-extension but not radial translation in abduction-adduction is the kinematic characteristic of the arthritic TM joint. We suggest that increased volar tilt is a predisposition to the dorsal translation of COR in the patient group. PMID- 15043899 TI - Palmar midcarpal instability: the results of treatment with 4-corner arthrodesis. AB - PURPOSE: Palmar midcarpal instability (PMCI) is one type of nondissociative carpal instability. The optimal treatment for PMCI is uncertain. The purpose of this investigation was to evaluate the results of capitate-lunate-triquetrum hamate (4-corner) arthrodesis for PMCI. METHODS: Over a 10-year period 8 patients were treated with 4-corner arthrodesis for PMCI. The instability was diagnosed clinically and confirmed with cineradiography. The symptomatic instability began in all patients after a notable trauma; 6 of the patients were injured at work and 2 were injured in a motor vehicle accident. Patients were evaluated at an average of 34 months after surgery. RESULTS: Seven of the 8 patients were satisfied with the surgery and 6 of 8 patients had no pain or mild pain. Both patients injured in motor vehicle accidents returned to all preinjury activities. Four of the 6 workers' compensation patients returned to work, 3 at full duty and 1 in a restricted position. Two patients, both with workers' compensation, did not return to their pre-injury jobs. One had occasional moderate pain and the other had chronic pain. The average wrist flexion/extension arc of motion decreased with surgery from 135 degrees to 75 degrees. The average grip strength increased significantly with surgery from 20 kg to 32 kg; however, grip strength remained significantly lower than the opposite side strength of 54 kg. CONCLUSIONS: Four-corner arthrodesis is a reasonable option for PMCI. Compared with previous reports of soft tissue reconstruction 4-corner arthrodesis may provide a more reliable solution for this difficult problem. PMID- 15043900 TI - Simultaneous radial closing wedge and ulnar shortening osteotomies for distal radius malunion. AB - PURPOSE: Closing wedge osteotomies are an attractive treatment option for distal radius malunion in patients with osteopenia; however, they require an ulnar head resection to accommodate closure of corrective osteotomy and to address the issue of ulnocarpal abutment. The literature contains little information on concomitant ulnar shortening osteotomy despite a physiologic solution. We report the functional and radiographic outcomes of 5 patients treated for symptomatic distal radius malunion with simultaneous radial closing wedge and ulnar shortening osteotomies. METHODS: All 5 patients were women aged 52 to 69 years (average, 61 years). Four patients had extra-articular radius fractures with dorsal angulation (20-22 degrees ) and shortening (3-7/mm); the other had the fracture with volar angulation (24 degrees ) and shortening (11 mm). Through a volar approach an appropriate amount of bone wedge was removed from the distal radius. A small volar T-plate was used to secure the osteotomized bone fragment. Six to 11 mm of ulnar shortening osteotomy was performed by using transverse osteotomy and compression plating technique with an AO compression device. RESULTS: In all 5 wrists healing of radial and ulnar osteotomies occurred less than 3 months after surgery. There were no postsurgical complications. Postsurgical radiographs showed that the volar tilt angle of the radius was reduced to normal range (range, 8-15 degrees ) in all wrists. The ulnar variance was 0 mm in 4 wrists and 2 mm in 1 wrist. There were significant improvements in pain, function, and range of motion at an average follow-up evaluation of 17 months. The average grip strength as a percentage of the opposite side improved from 30% before to 73% after surgery. CONCLUSIONS: This study showed that closing wedge osteotomy of the radius concomitant with ulnar shortening osteotomy is technically and functionally adequate. Our procedure is indicated for patients with osteopenia for whom opening wedge osteotomy of the radius is inadequate. PMID- 15043901 TI - Interfragmentary compression across a simulated scaphoid fracture--analysis of 3 screws. AB - PURPOSE: To measure the interfragmentary compression generated across a simulated fracture in cadaveric scaphoids by 3 different headless compression screws. METHODS: A transverse osteotomy was made through the waist of each scaphoid and a load cell to measure compression was interposed between the fragments, which were then fixed internally retrograde with either an Acutrak Standard (n = 10), Acutrak Mini (n = 12), or Bold (n = 10) screw. The surgeon was blinded to the measured compression, which was recorded during screw insertion and for the following 5 minutes. As a measure of scaphoid bone quality the modulus of elasticity of the trabecular bone of each specimen was then calculated from uniaxial compression tests. RESULTS: The mean interfragmentary compression generated by the Acutrak Standard screw was significantly greater and more consistent than the Bold screw or the Acutrak Mini screw. The compression achieved by the Acutrak Standard screw was also more consistent than that obtained by either the Bold or the Acutrak Mini screws as reflected by the lower standard deviation. The mean modulus of elasticity of the scaphoid trabecular bone was similar for each screw group. CONCLUSIONS: The interfragmentary compression generated by the Acutrak Standard screw was significantly greater and more consistent than that generated by either the Bold or Acutrak Mini screws. The compression generated by the Acutrak Standard and Mini screws was significantly better sustained over time than that generated by the Bold screw; however, these differences were small and may not be clinically important. The optimal compression required to promote scaphoid fracture union remains unknown and clinical trials are needed to further evaluate the outcome from using these devices. PMID- 15043902 TI - The role of the dorsal intercarpal ligament in dynamic and static scapholunate instability. AB - PURPOSE: Scapholunate instability (SLI) is the most common carpal instability. Recent studies have suggested that the dorsal intercarpal (DIC) and the dorsal radiocarpal ligaments play an important role in stabilization of the scaphoid and lunate. Differences between dynamic SLI and static SLI with a dorsal intercalated segment instability (DISI) are clearly described in the clinical literature; however, there has never been a clear explanation of the anatomic differences. This study describes the role of the DIC in the development of dynamic and static SLI with DISI in a cadaver model. METHODS: Five fresh cadavers were studied radiographically and by 3-dimensional digitization. Six increasing stages of instability were developed by sectioning progressively the following structures: the dorsal capsule, the palmar and proximal (membranous) portion of the scapholunate interosseous ligament, the DIC from its insertion on the scaphoid and trapezium, the dorsal scapholunate interosseous ligament from the scaphoid, the DIC ligament from its attachment on the lunate, and the lunotriquetral interosseous ligament. RESULTS: The scaphoid position and the scapholunate gap changed significantly after sectioning the entire scapholunate interosseous ligament and DIC from the scaphoid when a 5-kg load was applied. The lunate position was unchanged in both the loaded and the unloaded conditions. After detaching the DIC from the lunate, both the scaphoid and lunate moved and the scapholunate gap increased significantly in both loaded and unloaded conditions and showed a DISI deformity. CONCLUSIONS: This study describes an anatomic difference between dynamic and static scapholunate instability. Complete disruption of the scapholunate ligament did not result in the development of a static collapse of the lunate. The DIC had an important role in stabilizing the scaphoid and lunate and preventing DISI deformity. This study suggests that in the clinical setting the DIC ligament should be assessed intraoperatively and consideration should be given to repair and/or reconstruction of the DIC ligament attachments to both the scaphoid and the lunate. PMID- 15043903 TI - Wrist joint ganglion presenting as a painless mass in the palm: report of 2 cases. AB - Ganglions occur commonly in the wrist and arise from the radiocarpal and intercarpal joints. Although ganglions present commonly as masses on the dorsal or volar surface of the wrist, ganglions from wrist joints appear rarely at other locations in the hand. We report 2 cases of ganglions arising from wrist joints that presented as painless masses in the center of the palm without signs or symptoms of median or ulnar nerve compression. Surgical treatment required extensile exposure to trace the proximal stalks to their joints of origin. Knowledge of the possibility that a painless mass in the palm could be a ganglion arising from a joint in the wrist allows proper presurgical planning and informed consent. PMID- 15043904 TI - Passive strain distribution in the interosseous ligament of the forearm: implications for injury reconstruction. AB - PURPOSE: For severe forearm injuries such as an Essex-Lopresti fracture dislocation, functional reconstruction necessitates repair of the interosseous ligament (IOL) to restore normal load sharing between the radius and ulna. Locating or tensioning such a reconstruction improperly can lead to abnormal load sharing and/or restriction of forearm rotation. The normal IOL strains should indicate the proper location of reconstruction grafts and the proper forearm rotation for tensioning the grafts. The objective of this study was to quantify the passive strain distribution of the IOL of the forearm with passive rotation of the forearm throughout the range of motion. METHODS: The 3-dimensional motions of the radius with respect to the ulna were measured throughout forearm rotation in 10 cadaveric forearms by using an instrumented spatial linkage. From the bone motions and ligament insertion site geometry from dissection and computed tomographic scanning, insertion site motions were determined and used to calculate changes in ligament fiber lengths. RESULTS: The measured strain distribution in the IOL was nonuniform and varied with forearm rotation. The overall magnitude of IOL strain was found to be greatest in supination and smallest in pronation. In supination the strains varied across fibers with strains being greatest in the distal fibers and lowest in the proximal fibers. Strains in neutral rotation were uniform across fibers. Although fibers were generally slack in pronation proximal fibers were less slack than distal fibers. CONCLUSIONS: The results of this study indicate that fiber strains in the IOL vary from proximal to distal and depend on forearm rotation. Our data suggest that to prevent restriction of forearm rotation all grafts should be tensioned in supination, where measured strains were generally highest. Our data also suggest that a 2-bundle IOL reconstruction may be necessary for proper load transfer between the radius and ulna in both supination and pronation. PMID- 15043905 TI - Isolated compartment syndrome of the pronator quadratus compartment: a case report. AB - We present a case of compartment syndrome that involves the pronator quadratus space but does not affect any of the other forearm compartments. Symptoms consisted of pain out of proportion to examination findings, splinting of the forearm in a 30 degrees pronated position, and severe pain with passive supination. Diagnosis was confirmed with pressure measurements of all forearm compartments at the time of surgery. Symptoms resolved promptly with isolated release of the pronator quadratus compartment with concomitant reduction in compartment pressure in that one compartment. PMID- 15043906 TI - Limited, protected postsurgical motion does not affect the results of digital nerve repair. AB - PURPOSE: Casting for 3 to 4 weeks has been the accepted protocol after primary repair of digital nerve lacerations. In contrast, combined digital nerve and flexor tendon repairs are rehabilitated with immediate postsurgical range of motion. The purpose of this study was to compare the results of primary nerve repair in isolated digital nerve lacerations immobilized after surgery with nerve repairs combined with flexor tendon repairs that are mobilized in a limited, protected fashion immediately after surgery. METHODS: We reviewed retrospectively patients who had had surgical repair of isolated digital nerve lacerations or combined digital nerve and flexor tendon lacerations. Demographics recorded included age, hand dominance, injured digit, and time to mobilization. Follow-up data included range of motion at the metacarpophalangeal, proximal interphalangeal, distal interphalangeal, and wrist joints; static 2-point discrimination; and Semmes-Weinstein monofilament testing. Between-group comparisons were based on t-tests for continuous measures and chi-square tests for categoric measures. Paired t-tests were used for within-group comparisons. All comparisons were based on 2-tailed.05-level tests. RESULTS: Fourteen patients (16 digits) with isolated nerve repairs (group 1) and 12 patients (14 digits) with combined nerve and tendon repairs (group 2) were evaluated. The average age and duration at follow-up evaluation were similar in the 2 groups. The average time to mobilization, however, was 21 days in group 1 and 4 days in group 2. Injuries occurred equally in dominant and nondominant hands. Good range of motion returned in all digits. In addition there was no significant difference in final 2-point discrimination and Semmes-Weinstein testing between groups 1 and 2. CONCLUSIONS: Our data showed a decrease in sensibility between the injured and uninjured digits in each of the 2 groups studied, as has been shown previously. The difference in sensibility between the 2 groups, however, was not statistically significant. These data challenge the long-held belief that digital nerve repairs should be completely immobilized after surgery. PMID- 15043907 TI - Carpal tunnel syndrome: cross-sectional and outcome study in Ontario workers. AB - PURPOSE: To carry out an analytic cross-sectional study of Ontario workers with carpal tunnel syndrome (CTS) and to assess workers' symptoms, functional disabilities, recreational difficulties, and work capability 4 years after treatment of their CTS. METHODS: Data were obtained by review of Ontario Workers Safety and Insurance Board (WSIB) files and by completion of self-assessment questionnaires. Inclusion criteria included all workers registered with the Ontario WSIB who were off work with newly diagnosed carpal tunnel syndrome in 1996. RESULTS: There are 3 million workers covered by the WSIB in the province of Ontario. In 1996, 964 of them developed work-related CTS that required time off for treatment. Of these patients 53% were women and 75% had bilateral CTS. Eighty one percent of the unilateral cases involved the dominant extremity. The average age at the time of claim was 41 years and workers were at the same job type for an average of 7.4 years (unilateral) and 8.5 years (bilateral), respectively. Thirty-nine percent of workers had a history of another tendonitis or epicondylitis. Seventy-five percent of workers had surgery and on average returned to work 3 months later. Four years after treatment, outcome was assessed by self-administered questionnaires, for which there was a 73% response rate. Forty-six percent of workers experienced moderate to severe pain, 47% had moderate to severe numbness, and 40% had difficulty grasping and using small objects. Only 14% were symptom free. Successful return to work was considered to be a return to the same job with or without modifications, and it occurred in 64% of cases. Better clinical outcome scores were found to occur with surgery and abnormal nerve conduction study results. Worse clinical outcome scores were present with repeat surgery and surgical complications. Concurrent diagnoses of either tendonitis or epicondylitis also resulted in worse clinical outcome scores and worse return-to-work outcomes. The average total cost in Canadian dollars to the WSIB exceeded $13,700 per worker for a total cost in excess of $13,200,000 per year. (In 1996, $1 Canadian = $1.365 US.) CONCLUSIONS: These outcomes indicate that Canadian workers have a large amount of permanent pain and suffering, a large loss of work productivity, and incur a considerable financial cost as a result of work-related CTS. PMID- 15043908 TI - Measuring the strength of the intrinsic muscles of the hand in patients with ulnar and median nerve injuries: reliability of the Rotterdam Intrinsic Hand Myometer (RIHM). AB - PURPOSE: To determine the reliability and measurement error of measurements of intrinsic muscle strength of a new hand-held dynamometer (the Rotterdam Intrinsic Hand Myometer [RIHM]). METHODS: With the RIHM we obtained repeated measurements of the intrinsic muscle strength of the hand in 27 patients with peripheral nerve injury of the ulnar and/or median nerve in different stages of rehabilitation. The average time period after injury was 4.4 years (range, 99 days-11 years). RESULTS: Differences between 2 measurements greater than 6.3 N were interpreted as a real change in assessing the strength of the abduction of the little and index finger; for the median innervated muscles of the thumb this value was 16 N. CONCLUSIONS: In patients with nerve injuries the muscle strength is usually assessed with manual muscle strength testing and grip- and pinch-strength dynamometers. Preferably the intrinsic muscle strength should be measured in isolation and quantitatively. The RIHM is a new dynamometer that allows for measurements of the intrinsic muscle strength in isolation with reliability comparable to grip and pinch measurements. PMID- 15043909 TI - The V-sling: a modified medial intermuscular septal sling for anterior transposition of the ulnar nerve. AB - Subcutaneous anterior ulnar nerve transposition has been advocated by many surgeons as simple and effective. Techniques to maintain the nerve anterior to the medial epicondyle include subcutaneous pocket, subcutaneous-fascia tunnel, and fascial and fasciodermal sling. We describe a modified technique that uses the medial intermuscular septum as a sling to allow a more gentle transition of the ulnar nerve as it enters into the flexor carpi ulnaris muscle belly. PMID- 15043911 TI - Redefining the "arcade of Struthers". PMID- 15043910 TI - Interobserver and intraobserver reliability of therapist-assisted videotaped evaluations of upper-limb hemiplegia. AB - PURPOSE: Therapist-assisted videotaped sessions have been used to augment physical examinations in the evaluation of hand and arm function in patients with spastic hemiplegia. The purpose of this study was to assess the interobserver and intraobserver reliability of standardized videotaped examinations in the evaluation and functional classification of these patients. METHODS: Three examiners reviewed standardized videotaped examinations of 10 adolescents with spastic hemiplegia on 2 separate occasions. All 10 patients were under consideration for surgical intervention for their upper-extremity dysfunction. Videotapes were used to assess upper-extremity range of motion, finger and thumb deformity, and reach, pinch, and grip function. Upper-extremity function was graded according to the House and Mowery classification systems. Interobserver and intraobserver reliabilities were measured with the kappa coefficient. RESULTS: Range of motion, deformity, and upper-extremity functional strategy assessment showed slight to excellent interobserver reliability and good to almost perfect intraobserver reliability. Interobserver and intraobserver reliability of the consolidated House classification system was more reliable than the Mowery or standard House classification systems. CONCLUSIONS: Evaluations of standardized videotaped examinations in patients with hemiplegia were reliable between and among observers. Such therapist-assisted videotaped evaluations may provide useful data for clinical decision-making and multicenter outcomes studies in patients with upper-extremity involvement with spastic hemiplegia. PMID- 15043916 TI - Sedimentation equilibrium and the foundations of protein chemistry. PMID- 15043917 TI - Those wonderful early years with the Model E ultracentrifuge and David Yphantis. PMID- 15043918 TI - Graduate student days at MIT. PMID- 15043919 TI - Spinning with Dave: David Yphantis's contributions to ultracentrifugation. AB - For nearly 50 years David Yphantis has helped advance analytical ultracentrifugation, promoted rigor in the thermodynamic analysis of biochemical data and encouraged students and colleagues to look for the deepest possible understanding of science. This article, written by five of Dave's students, presents some of the impressions he has made over the years. PMID- 15043920 TI - Thermodynamics of glycophorin A transmembrane helix dimerization in C14 betaine micelles. AB - We have used sedimentation equilibrium analytical ultracentrifugation to measure the free energy change for the glycophorin A transmembrane helix-helix dimerization in C14 betaine micelles. By varying the amount of micellar C14 betaine, we show that the protein association reaction in the micellar C14 phase behaves as an ideal-dilute solution. In this hydrophobic environment, the mole fraction standard state free energy change for self-association of the SNGpA99 glycophorin A construct is -5.7 (+/-0.3, N=5) kcal mol(-1) at 25 degrees C. Compared with previous results carried out in C(8)E(5) micellar solutions, the free energy of dimerization is 1.3 kcal mol(-1) less favorable in C14 betaine micelles. In contrast, when considered on a per-interface basis, the formation of the glycophorin A transmembrane dimer in C14 betaine micelles may be more favorable than the association of several designed transmembrane peptides. PMID- 15043921 TI - Nonequilibrium thermodynamics of membrane-confined electrophoresis. AB - Membrane-confined electrophoresis (MCE) is an electrophoretic transport method in which macromolecules in solution are confined within a cuvette through which a current flows. Small ions that can permeate the membranes permit current flow. The method is the electrophoretic analog to analytical ultracentrifugation. Systems in the MCE instrument are described by nonequilibrium thermodynamics. This description forms the basis of a program, implemented using finite element methods, that can model transport processes in such systems over an extended time, from arbitrary starting conditions to steady state. Issues relevant to the analysis of systems in which macromolecular species are involved in mass-action associations are discussed. Particular attention is given to steady-state electrophoresis, from which measurements of reduced molecular charge are sought. The relationship of such measurements to valence is discussed. PMID- 15043922 TI - Re-examining the oligomerization state of macrophage migration inhibitory factor (MIF) in solution. AB - The state of oligomerization of macrophage migration inhibitory factor (MIF, also known as glycosylation inhibiting factor, GIF) in solution has been variously reported as monomer, dimer, trimer, or mixtures of all three. Several crystal structures show MIF to be a trimer. Sedimentation velocity shows a recombinant human MIF sample is quite homogeneous, with 98% as a species with s(20,w)=3.07 S and D(20,w)=8.29 x 10(-7) cm(2)/s. Using the partial specific volume calculated from the amino acid composition these values imply a mass of 33.56 kDa, well above that of dimer, but also 9% below the trimer mass of 37.035 kDa. Sedimentation equilibrium data at loading concentrations from 0.01 to 1 mg/ml show unequivocally that the self-association is extremely tight. However, the apparent mass is 33.53 kDa [95% confidence 33.25-33.82], again 9% below that expected for 100% trimer. To examine the possibility this protein has an unusual partial specific volume, sedimentation equilibrium was also done in H(2)O/D(2)O mixtures, giving 0.765+/-0.017 ml/g rather than the calculated 0.735 ml/g. With this revised partial specific volume, the equilibrium and velocity data each give M=37.9+/-2.8 kDa, fully consistent with a strongly-associated trimeric quaternary structure. PMID- 15043923 TI - Sedimentation equilibrium in a solution containing an arbitrary number of solute species at arbitrary concentrations: theory and application to concentrated solutions of ribonuclease. AB - Simple expressions are derived describing the equilibrium concentration gradient of each species in a solution containing an arbitrary number of solute species at arbitrary concentration, as a function of the concentration of all species. Quantitative relationships between the species gradients and experimentally observable signal gradients are presented. The expressions are model-free and take into account both attractive and repulsive interactions between all species. In order to analyze data obtained from strongly nonideal solutions, a statistical thermodynamic model for repulsive solute-solute interactions is required. The relations obtained are utilized to analyze the dependence of the equilibrium gradient of ribonuclease A in phosphate-buffered saline, pH 7.4, upon total protein concentration. Experimental results are interpreted in the context of a model for weak self-association leading to the formation of significant amounts of oligomers at total protein concentrations exceeding 25 g/l. PMID- 15043924 TI - HIV Rev self-assembly is linked to a molten-globule to compact structural transition. AB - By regulating the differential expression of proviral pre mRNA in the host cell, Rev plays a crucial role in the HIV-1 life cycle. The capacity of Rev to function is intimately linked to its ability to self-associate. Nevertheless, little is known about the exact role of self-association in the molecular mechanism defining its biological activity. A prerequisite knowledge is a definition of the molecular events undertaken by Rev during the process of self-assembly. Thus, this study was initiated to monitor the structure of Rev as a function of protein concentration. Rev undergoes a structural transition as a consequence of self assembly. This structural transition was monitored by three spectroscopic methods. The accessibility of the single tryptophan in Rev monomer to acrylamide quenching increases with decreasing protein concentration. At very low concentration of Rev, the tryptophan accessibility is close to that of an unfolded Rev. As evaluated by circular dichroism, the secondary structure of Rev is protein concentration dependent as evidenced by an increase in the magnitude of ellipticity with increasing protein concentration. Further, results from ANS binding studies indicate that the ANS binding sites in Rev experience an apparent increase in hydrophobicity as the Rev concentration was increased. These concentration dependent changes seem to reach a maximum above 5 microM Rev monomer concentration. In order to define the mode of Rev self-association sedimentation velocity and equilibrium experiments were conducted. There are evidently two consecutive progressive association processes. At protein concentrations below 0.5 mg/ml, the data from sedimentation studies can be fitted to a single isodesmic model. Simulation of velocity sedimentation profile indicates that free Rev monomer that has not entered into the association processes can best be described to exhibit a value of S(20,w) that is substantially smaller than 1.4 S, a value needed to fit the rest of the data. The latter value is consistent for a Rev monomer with the expected molecules weight and if it were to assume a compact globular shape. These spectroscopic and hydrodynamic results imply that monomeric Rev is in a molten globule state, which becomes more compact upon self-association. PMID- 15043925 TI - Reverse smoothing: a model-free data smoothing algorithm. AB - Biophysical chemistry experiments, such as sedimentation-equilibrium analyses, require computational techniques to reduce the effects of random errors of the measurement process. The existing approaches have primarily relied on assumption of polynomial models and least-squares approximation. Such models by constraining the data to remove random fluctuations may distort the data and cause loss of information. The better the removal of random errors the greater is the likely introduction of systematic errors through the constraining fit itself. An alternative technique, reverse smoothing, is suggested that makes use of a more model-free approach of exponential smoothing of the first derivative. Exponential smoothing approaches have been generally unsatisfactory because they introduce significant data lag. The approaches given here compensates for the lag defect and appears promising for the smoothing of many experimental data sequences, including the macromolecular concentration data generated by sedimentation equilibria experiments. Test results on simulated sedimentation-equilibrium data indicate that a 4-fold reduction in error may be typical over standard analyses techniques. PMID- 15043926 TI - Global analysis of non-specific protein-nucleic interactions by sedimentation equilibrium. AB - Protein-nucleic acid interactions govern a variety of processes, including replication, transcription, recombination and repair. These interactions take place in both sequence-specific and non-specific modes, and the latter occur in many biologically significant contexts. Analytical ultracentrifugation is a useful method for the detailed characterization of the stoichiometry and affinity of macromolecular interactions in free solution. There has been a resurgence of interest in the application of sedimentation equilibrium methods to protein nucleic acid interactions. However, these studies have been generally focused on sequence-specific interactions. Here we describe an approach to analyze non specific interactions using sedimentation equilibrium. We have adapted an existing model for non-specific interaction of proteins with finite, one dimensional nucleic acid lattices for global fitting of multiwavelength sedimentation equilibrium data. The model is extended to accommodate protein binding to multiple faces of the nucleic acid, resulting in overlap of consecutive ligands along the sequence of the RNA or DNA. The approach is illustrated in a sedimentation equilibrium analysis of the interaction of the double-stranded RNA binding motif of protein kinase R with a 20-basepair RNA construct. PMID- 15043927 TI - Characterization of macromolecular heterogeneity by equilibrium sedimentation techniques. AB - New graphical procedures have been developed to investigate the heterogeneity of protein preparations using sedimentation equilibrium. The heterogeneous systems that can be studied include self-associating systems contaminated by incompetent monomer, self-associating systems contaminated by non-dissociating oligomer and simple non-interacting monomer-oligmer disperse systems. The new procedures are based on the concentration dependence of the apparent association constants estimated by a non-linear least square fitting program (NONLIN), on the assumption of conservation of mass during sedimentation and on the applications of several standard techniques for statistical inferences of NONLIN estimations. The procedures outlined here can detect various types of heterogeneity, discriminate amongst different types of heterogeneity, estimate the amount of contaminant causing heterogeneity and determine the true equilibrium constant of the self-associating components. The procedures appear to be sensitive, accurate and easily applicable when tested using both protein samples and computer simulated data. PMID- 15043928 TI - Fluorescence detection for the XLI analytical ultracentrifuge. AB - Analytical ultracentrifugation (AUC) provides first-principle hydrodynamic and thermodynamic information concerning the size, shape and interactions of macromolecules. The fundamental measurement needed in AUC is the macromolecular concentration as a function of radial position and time. Currently, the Beckman Coulter XLI analytical ultracentrifuge may be equipped with absorbance and refractive detectors, which provide complementary concentration determinations. For detecting trace quantities of materials, fluorescence detection offers unique advantages over either absorbance or interference detection. A prototype fluorescence detector for the XLI analytical ultracentrifuge has been developed and its characteristics determined. An Ar(+) laser provides a continuous 488-nm excitation beam. Radial resolution is achieved by scanning the focused beam along a radial axis. Detection of the fluorescence signal uses a co-axial, front-face optical configuration to reduce inaccuracies in the concentration caused by inner filter effects. A high-speed A/D data acquisition system allows the fluorescence intensity to be monitored continuously and at a sufficiently high angular resolution so that at any radial position the intensities from all of the samples may be acquired at each revolution. The fluorescence detector is capable of detecting concentrations as low as 300 pM for fluorescein-like labels. The radial resolution of the fluorescence detector is comparable to that of the absorbance system. Both sedimentation velocity and sedimentation equilibrium measurements may be made with the fluorescence detector. Results are presented comparing data acquired using the fluorescence with those acquired using the absorbance detector. PMID- 15043929 TI - A model for sedimentation in inhomogeneous media. I. Dynamic density gradients from sedimenting co-solutes. AB - Macromolecular sedimentation in inhomogeneous media is of great practical importance. Dynamic density gradients have a long tradition in analytical ultracentrifugation, and are frequently used in preparative ultracentrifugation. In this paper, a new theoretical model for sedimentation in inhomogeneous media is presented, based on finite element solutions of the Lamm equation with spatial and temporal variation of the local solvent density and viscosity. It is applied to macromolecular sedimentation in the presence of a dynamic density gradient formed by the sedimentation of a co-solute at high concentration. It is implemented in the software SEDFIT for the analysis of experimental macromolecular concentration distributions. The model agrees well with the measured sedimentation profiles of a protein in a dynamic cesium chloride gradient, and may provide a measure for the effects of hydration or preferential solvation parameters. General features of protein sedimentation in dynamic density gradients are described. PMID- 15043930 TI - A model for sedimentation in inhomogeneous media. II. Compressibility of aqueous and organic solvents. AB - The effects of solvent compressibility on the sedimentation behavior of macromolecules as observed in analytical ultracentrifugation are examined. Expressions for the density and pressure distributions in the solution column are derived and combined with the finite element solution of the Lamm equation in inhomogeneous media to predict the macromolecular concentration distributions under different conditions. Independently, analytical expressions are derived for the sedimentation of non-diffusing particles in the limit of low compressibility. Both models are quantitatively consistent and predict solvent compressibility to result in a reduction of the sedimentation rate along the solution column and a continuous accumulation of solutes in the plateau region. For both organic and aqueous solvents, the calculated deviations from the sedimentation in incompressible media can be very large and substantially above the measurement error. Assuming conventional configurations used for sedimentation velocity experiments in analytical ultracentrifugation, neglect of the compressibility of water leads to systematic errors underestimating sedimentation coefficients by approximately 1% at a rotor speeds of 45000 rpm, but increasing to 2-5% with increasing rotor speeds and decreasing macromolecular size. The proposed finite element solution of the Lamm equation can be used to take solvent compressibility quantitatively into account in direct boundary models for discrete species, sedimentation coefficient distributions or molar mass distributions. Using the analytical expressions for the sedimentation of non-diffusing particles, the ls g*(s) distribution of apparent sedimentation coefficients is extended to the analysis of sedimentation in compressible solvents. The consideration of solvent compressibility is highly relevant not only when using organic solvents, but also in aqueous solvents when precise sedimentation coefficients are needed, for example, for hydrodynamic modeling. PMID- 15043931 TI - A comparison of weight average and direct boundary fitting of sedimentation velocity data for indefinite polymerizing systems. AB - Analysis of sedimentation velocity data for indefinite self-associating systems is often achieved by fitting of weight average sedimentation coefficients (s(20,w)) However, this method discriminates poorly between alternative models of association and is biased by the presence of inactive monomers and irreversible aggregates. Therefore, a more robust method for extracting the binding constants for indefinite self-associating systems has been developed. This approach utilizes a set of fitting routines (SedAnal) that perform global non-linear least squares fits of up to 10 sedimentation velocity experiments, corresponding to different loading concentrations, by a combination of finite element simulations and a fitting algorithm that uses a simplex convergence routine to search parameter space. Indefinite self-association is analyzed with the software program isodesfitter, which incorporates user provided functions for sedimentation coefficients as a function of the degree of polymerization for spherical, linear and helical polymer models. The computer program hydro was used to generate the sedimentation coefficient values for the linear and helical polymer assembly mechanisms. Since this curve fitting method directly fits the shape of the sedimenting boundary, it is in principle very sensitive to alternative models and the presence of species not participating in the reaction. This approach is compared with traditional fitting of weight average data and applied to the initial stages of Mg(2+)-induced tubulin self-associating into small curved polymers, and vinblastine-induced tubulin spiral formation. The appropriate use and limitations of the methods are discussed. PMID- 15043932 TI - Analysis of heterologous interacting systems by sedimentation velocity: curve fitting algorithms for estimation of sedimentation coefficients, equilibrium and kinetic constants. AB - Analytical ultracentrifugation (AUC) has played and will continue to play an important role in the investigation of protein-protein, protein-DNA and protein ligand interactions. A major advantage of AUC over other methods is that it allows the analysis of systems free in solution in nearly any buffer without worry about spurious interactions with a supporting matrix. Large amounts of high quality data can be acquired in relatively short times. Advances in software for the treatment of AUC data over the last decade have eliminated many of the tedious aspects of AUC data analysis, allowing relatively rapid analysis of complicated systems that were previously unapproachable. A software package called sedanal is described that can perform global fits to AUC sedimentation velocity data obtained for both interacting and non-interacting, macromolecular multi-species, multi-component systems, by combining data from multiple runs over a range of sample concentrations and component ratios. Interaction parameters include both forward and reverse rate constants, or equilibrium constants, for each reaction, as well as concentration dependence of both sedimentation and diffusion coefficients. sedanal fits to time-difference data to eliminate time independent systematic errors inherent in AUC data. The sedanal software package is based on the use of finite-element numerical solutions of the Lamm equation. PMID- 15043933 TI - NOVOcan: a molecular link among selected glial cells. AB - The nervous system is generated from cells lining the ventricular system. Our understanding of the fate potentials and lineage relationships of these cells is being re-evaluated, both because of recent demonstrations that radial glia can generate neurons and because of the identification of fate-determining genes. A variety of intrinsic and extrinsic molecules, including proteoglycans, regulate embryonic and postnatal brain development. Using probes modeled after species conserved domains of heparan sulfate proteoglycans, we cloned a novel gene called novocan, raised monoclonal antibodies against a segment of the predicted amino acid sequence of the expressed protein (NOVOcan) and used the antibodies to establish the cell and tissue localization of NOVOcan in postnatal rat brains by immunohistochemistry. NOVOcan was expressed in cells lining the ventricles, including a variety of radial glia during early postnatal development. Later, as radial glia disappeared and ependymal cells appeared, NOVOcan was detected in ependymal cells and in tanycytes, a specialized form of ependymal cell resembling radial glia. NOVOcan was absent in two known progeny of radial glia, mature astrocytes and neurons. Whereas NOVOcan was also absent in mature oligodendrocytes (OLGs), it was present in OLG precursors in developing white matter. These studies set the stage for determining the roles of NOVOcan in brain cell lineage patterns as well as in other aspects of development. PMID- 15043934 TI - Hydrodynamic bead modelling of the 2:1 p50-IkappaBgamma complex. AB - NFkappaB is an important and ubiquitous transcription factor formed by various homo- and heterodimers of the NFkappaB family. The active transcription factor regulates genes involved in immune, inflammatory and survival responses. Specificity in gene regulation is achieved, at least in part, by the distinct DNA binding preferences of the various homo- and heterodimers and by the complex pathways that lead to signal-induced degradation of the IkappaB inhibitors. Analytical ultracentrifugation and hydrodynamic bead modelling were used to model the solution structures of the NFkappaB family member p50, its inhibitor IkappaBgamma and their complex. Sedimentation equilibrium (SE) and sedimentation velocity (SV) data show that p50 is a dimer in solution with a sedimentation coefficient consistent with a conformation intermediate between the closed conformation observed in the crystal structure of the p50 (N-terminal domain)-p65 heterodimer complexed with IkappaBalpha and the open conformation adopted by p50 when bound to DNA. SE and SV data show that IkappaBgamma is a monomer in solution and is prone to aggregation over time. p50 forms a 2:1 stoichiometric complex with IkappaBgamma in solution with a sedimentation coefficient consistent with a closed conformation for the p50 dimer. PMID- 15043935 TI - Sedimentation velocity, multi-speed method for analyzing polydisperse solutions. AB - A method is described for the sedimentation velocity analysis of solutions composed of macromolecular solutes of widely disparate size. In sedimentation velocity experiments, usually a single rotor speed is chosen for the entire run, and consequently, the range of observable sedimentation coefficients can be severely limited. This limitation can be removed if the speed is varied during the run, starting with a relatively low speed so that the largest particles can be easily observed. The speed is increased during the run until full speed is attained and the run continued at full speed until the smallest species of interest have cleared the solution. The method, called wide distribution analysis, is based on the method developed originally by Yphantis and co-workers (Proc. Natl. Acad. Sci. USA 78 (1981) 1431) and on the time derivative method of Stafford (Anal. Biochem. 203 (1992) 295), essentially eliminating both the time independent and radially-independent noise thereby improving the precision, especially for interference optics. An algorithm for analysis of data from both absorbance and interference optics and experimental protocols compatible with the Beckman XL-I Analytical Ultracentrifuge are presented. With these protocols an extremely wide range of sedimentation coefficients from approximately 1.0 to 250000 S can be accommodated in a single multi-speed run. PMID- 15043936 TI - Cloning and expression of the AID gene in the channel catfish. AB - A full-length activation-induced cytidine deaminase (AID or Aicda) cDNA has been obtained from the channel catfish (Ictalurus punctatus). A single open reading frame predicts a 209 amino acid protein that has 57% identity and 73% similarity with the AID proteins of mouse and human. All residues that have previously been found to be critical for deamination, as well as for somatic hypermutation, are conserved in the catfish AID. These residues are also conserved in AID proteins predicted, from genome database sequences, to be expressed in Fugu and zebrafish. The catfish AID is expressed at low levels in spleen, kidney, intestine and fin margins, but not in muscle, liver or brain. Immunoglobulin heavy chain (IgH) is also expressed in the tissues where AID is expressed. The 'ectopic' expression of AID in non-lymphoid tissue was unexpected and not readily explained. However, the identification of a fish AID gene will allow us to determine the tissue architecture and locations for affinity maturation in fish. PMID- 15043937 TI - The ratio of full length IgY to truncated IgY in immune complexes affects macrophage phagocytosis and the acute phase response of mallard ducks (Anas platyrhynchos). AB - Ducks produce a full length IgY and a truncated isoform (IgYDeltaFc). IgY and IgY(DeltaFc) were isolated from ducks vaccinated against Escherichia coli and used to make E.coli-Ig immune complexes (IC). Phagocytosis of IC by duck monocytes decreased directly with the proportion of IgYDeltaFc (p<0.001). IC containing IgY:IgY(DeltaFc) at ratios of 100:0, 50:50, 0:100, and 0:0 (E. coli alone) were injected intravenously into naive mallard ducks. At 24 h after injection, plasma hemopexin levels were higher in ducks given either the 0:100 ratio or the 100:0 ratio than those given 50:50 or E. coli alone (p<0.005) Liver IL-1beta mRNA levels followed a similar pattern. Splenic IL-1beta mRNA decreased markedly as the proportion of IgY(DeltaFc) increased (p<0.01) Thus, IgY(DeltaFc) may shift the response to IC from the spleen to the liver as infections progress from acute to chronic. PMID- 15043938 TI - Recent findings on phenoloxidase activity and antimicrobial activity of hemocyanins. PMID- 15043939 TI - Microbial infection causes the appearance of hemocytes with extreme spreading ability in monolayers of the tobacco hornworm Manduca sexta. AB - The ability to adhere to and spread on a surface is a common property of insect blood cells. Spreading on a glass surface by insect hemocytes is often used as a measure of immune fitness that can be inhibited by some insect pathogens and parasites. Here, we report that upon infection of the tobacco hornworm Manduca sexta with either a fungus (Beauveria bassiana) or a bacterium (Photorhabdus luminescens), a new type of hemocyte, not previously observed in healthy insects, was found in hemocyte monolayers. These cells have a distinctive morphology, characterised by extreme spreading ability. They achieve a diameter of up to 120 microm after 1 h on glass coverslips and are therefore extremely thin. These hyper-spreading cells first appear in fungal-infected insects prior to hyphal growth. Their numbers later fall to zero as the pathogen begins to proliferate. The same hyper-spreading cells are induced after a 24 h delay following an injection of laminarin, a source of the fungal cell wall polymer beta-1,3 glucans. Wounding, on the other hand, did not cause the appearance of hyper spreading cells. Evidence is presented here that is consistent with these spreading cells having a role in the cellular immune response of nodule formation. PMID- 15043940 TI - Effect of experimental microbial challenge on the expression of defense molecules in Eisenia foetida earthworm. AB - Earthworms are able to protect themselves against invading pathogens due to efficient innate defense mechanisms. Currently, two types of antimicrobial factors including lysozyme-like molecule and factors with hemolytic activity, as well as a pattern recognition protein named coelomic cytolytic factor (CCF) have been identified in Eisenia foetida earthworms. However, the modulations of these defense molecules during in vivo immune response have not been addressed. In this study, we investigated the effect of experimental challenge with live Gram negative and Gram-positive bacteria and with beta-1,3-glucan on the expression of CCF and the hemolytic factor fetidin. In parallel, we followed levels of hemolytic activity and lysozyme-like activity in the coelomic fluid of challenged earthworms. We show that the biosynthesis of CCF, but not fetidin, is up regulated upon microbial stimulation. Parenteral administration of bacteria or microbial polysaccharides in earthworms results, in the coelomic fluid, in augmented level of CCF, increased lysozyme-like activity and decreased hemolytic activity. The decreased hemolytic activity of the coelomic fluid reflects the increase of the whole protein content in the absence of synthesis of hemolytic proteins. PMID- 15043941 TI - Molecular cloning of a beta-glucan pattern-recognition lipoprotein from the white shrimp Penaeus (Litopenaeus) vannamei: correlations between the deduced amino acid sequence and the native protein structure. AB - The hemolymph pattern-recognition beta-glucan binding protein from the white shrimp Penaeus (Litopenaeus) vannamei is also a high density lipoprotein (betaGBP HDL) involved in innate immunity. The betaGBP-HDL full length cDNA sequence determined was 6.3 kb long, and contains a long 3'UTR region with a polyadenylation signal and a poly-A+ tail. The open reading frame is 1454 amino acids long and the N-terminal residue of the mature protein is localized in position 198 of the ORF. Comparison of the betaGBP-HDL amino acid sequence against GenBank detected only significant similarity to betaGBP from the crayfish Pacifastacus leniusculus. betaGBP-HDL is expressed in hepatopancreas, muscle, pleopods and gills, but not in hemocytes as determined by RT-PCR. We discuss the analysis of the deduced primary sequence in terms of the predicted secondary structure, glucanase-like and RGD motives relevant to its dual roles in defence and lipid transport. PMID- 15043942 TI - Differentially expressed genes that encode potential markers of goldfish macrophage development in vitro. AB - Primary kidney macrophage (PKM) cultures derived from goldfish hematopoietic tissues develop from early progenitors to mature macrophages in response to endogenous growth factor(s). When grown in vitro, PKM shift from a proliferative phase, where most of the proliferation and differentiation events take place, to a senescence phase, where there is cessation of proliferation and differentiation events and ultimately cell death through a process of apoptosis. The phenotypic changes of PKM from the proliferative to senescence phase are a reflection of specific changes in gene expression; therefore, comparison of gene expression patterns between the two phases should lead to the identification of macrophage genes directly involved in the positive and negative regulation of hematopoietic events, as well as genes that are modulated downstream from these regulatory points. Differential cross-screening of the proliferative phase PKM cDNA library using proliferative and senescence phase (32)P-labeled cDNA probes identified several differentially expressed genes. Specifically, initial screen of 9200 clones yielded 734 differential primary clones that were isolated and analyzed using a PCR-based secondary screen. The majority of these clone isolates encoded a single transcript as determined by PCR amplification of the primary clones. The secondary screen confirmed the differential expression of 306 clones (3.32% of the total number of screened clones). Two hundred and forty four clones were sequenced; 158 and 86 were preferentially expressed during proliferative and senescence phases, respectively. Several potential candidates of fish macrophage hematopoiesis were identified. These include, for example, zinc finger protein 147, nucleophosmin, 14-3-3 protein, adenine nucleotide translocator 2 (ANT2), granulin, survivin-1, and apoptosis inhibitor-5. In addition, several potential markers of macrophage differentiation and/or function were identified and their expression patterns characterized across three distinct stages of macrophage development in vitro. These include legumain, CD63, interferon-inducible protein, macrosialin (CD68), transcription factor MafB, and the molecular chaperone BiP/GRP78. These analyses will facilitate future characterization of macrophage developmental events by providing a more global perspective of various facets of macrophage hematopoiesis. PMID- 15043943 TI - Organization and expression analysis of the zebrafish hepcidin gene, an antimicrobial peptide gene conserved among vertebrates. AB - Hepcidin is an antimicrobial peptide and iron-regulatory molecule that is conserved among vertebrates. Mutations or over-expression of the human hepcidin gene have been found in patients with hemochromatosis and refractory anemia. To further understand the function and regulation of hepcidin, animal models are needed. We sequenced cDNA, genes and upstream regions of zebrafish hepcidin and analyzed gene expression by kinetic PCR. Zebrafish hepcidin genes consist of two introns and three exons that encode a prepropeptide (91 amino acids). The amino acid sequences and gene organization were remarkably conserved between zebrafish and other species. Elevated gene expression was observed in abdominal organs, skin, and heart in fish that developed signs of infection following bacterial injection. Zebrafish may be a suitable model organism for further study of hepcidin gene regulation. PMID- 15043944 TI - T cells and the thymus in developing zebrafish. AB - The expression of genes encoding T cell receptor (TCR) alpha was used to follow the development of the thymus and to analyze the distribution of T cells in zebrafish. In the thymus, expression was first detected, by in situ hybridization, at four days post fertilization. In RNA extracted from whole fish, TCRalpha transcripts were also detected at four days and reached adult levels at three weeks. At six weeks, TCRalpha was expressed throughout the thymus, whereas rag1 expression was localized to the peripheral regions. Expression of TCRalpha outside the thymus was detected at nine days. In adult peripheral organs, the greatest expression was in the pronephros, mesonephros and intestine; expression in the spleen became greater as fish age. Three new, relatively highly expressed, TCR Valpha families were identified. PMID- 15043945 TI - Sequence analysis and expression of a CXC chemokine in resistant and susceptible catfish after infection of Edwardsiella ictaluri. AB - Chemokines represent a superfamily of chemotactic cytokines involved in recruitment, activation and adhesion of a variety of leukocyte types to inflammatory foci. We cloned and sequenced the cDNA of a CXC chemokine that is most similar to CXCL10 from channel catfish and blue catfish. Sequence analysis of PCR amplicons from a single F1 hybrid catfish indicated that channel catfish and blue catfish may have a multigene family for the CXC chemokine. The catfish CXC chemokine was expressed in a wide range of tissues including head kidney, spleen, liver, gill, skin, stomach, and intestine, but not in the muscle. Fish challenged with intracellular bacterium Edwardsiella ictaluri, the causative agent of enteric septicemia of catfish (ESC), showed dramatically elevated levels of the CXC chemokine expression, as quantified with real time RT-PCR. Differential expression profiles were observed between resistant and susceptible channel catfish strains and blue catfish. Blue catfish were characterized by only modest induction in comparison to the drastic elevation of the CXC chemokine in channel catfish. PMID- 15043946 TI - Allelic polymorphism of TCRalpha chain constant domain genes in the bicolor damselfish. AB - T cell receptor (TCR) chains are composed of two extracellular domains, the membrane-distal variable domain and the membrane-proximal constant domain. Data presented here show that the TCRA 'constant' (C) domain of damselfish exhibits considerable allelic polymorphisms that appear to be positively selected. Each of 32 damselfish TCRAC clones showed different patterns of atypical polymorphism in the constant region. Twenty-three of the 121 TCRalpha constant region amino acid residues show substitutions, clustered mainly in the loops between the beta strands. Coding regions of the TCRAC genes differ by up to 8% at the nucleotide level and 20% at the amino acid level. Southern hybridization, polymorphism segregation, and genomic cloning data suggest allelic polymorphism at two TCRAC genes, distinguished by a single amino acid. KA/KS ratios suggest that balancing selection is acting to maintain polymorphisms at the variable sites of one of these genes, but not the other, in a manner comparable to the peptide binding regions of MHC. Nonetheless, each TCRAC gene is spliced to variable and joining segments similar to those described in other species. These data suggest that our understanding of the function of the TCR constant domains of these vertebrates is incomplete. PMID- 15043947 TI - An Mx1 promoter-reporter system to study interferon pathways in rainbow trout. AB - A rainbow trout interferon (IFN) reporter system has been established by selection of a stable cell line, RTG-P1, transfected with a plasmid expressing the firefly luciferase gene under the control of the promoter for the IFN-induced gene Mx1. After 148 passages, the luciferase expression was still highly induced by polyinosinic:polycytidylic acid (poly I:C) in RTG-P1 cells. Different IFN inducers (dsRNA, viral hemorrhagic septicaemia virus or conditioned medium containing rainbow trout antiviral activity) were able to stimulate the IFN reporter system in RTG-P1, showing that this cell line can be used to study the activation of the IFN pathway in various contexts. Pyrrolidine dithiocarbamate (PDTC), an NF-kappaB inhibitor, significantly blocked poly I:C induced luciferase accumulation in RTG-P1 at intermediate doses (1-10 microM), suggesting that Mx1 induction through the IFN signalling pathway is NF-kappaB-dependent in fish. This inhibition was not observed for doses of 50 microM or higher. The RTG-P1 reporter system constitutes an interesting tool to study the induction and regulation of IFN signalling in teleost fish. PMID- 15043948 TI - Use of staphylococcal protein A in the analysis of teleost immunoglobulin structural diversity. AB - Staphylcoccal protein A (SpA) adsorption and sephacryl-S300 filtration were employed to isolate Ig from the sera of six aquaculturally important teleost species; Morone saxatilis (striped bass), Lates calcarifer (barramundi), Oreochromis mossambicus (Mozambique tilapia) and Oreochromis niloticus (Nile tilapia), Salmo salar (Atlantic salmon), and Oncorhynchus mykiss (rainbow trout). While both gel filtration (S300) and SpA adsorption could purify the 800 kDa tetrameric Ig, SpA demonstrated species-specific variability in the amount retrieved. Virtually 100% of this high molecular weight Ig could be isolated from Mosambique tilapia serum, while 84, 17, 10.7 and 0.5% could be isolated from barramundi, striped bass, Nile tilapia, and Atlantic salmon, respectively. Significant amounts of Ig could not be isolated (<0.1%) from rainbow trout (O. mykiss) serum. All SpA-isolated proteins were approximately 800 kDa in molecular weight and were solely composed of equimolar concentrations of H ( approximately 75 kDa) and L ( approximately 25 kDa) chains. Denaturing, non-reducing electrophoresis demonstrated differential disulfide polymerization (redox forms) consistent with those observed with other teleost species; however, SpA exhibited less affinity for Ig possessing completely polymerized tetramers than the more reduced forms, with the exception of Mossambique tilapia. The existence of three different molecular weight H chains (75, 85, 95 kDa) in Nile tilapia was also observed. Each redox form of Nile tilapia Ig incorporated only one size of H chain. PMID- 15043949 TI - Generation, purification and functional characterization of three C3a anaphylatoxins in rainbow trout: role in leukocyte chemotaxis and respiratory burst. AB - Activation of the complement system leads to cleavage of the C3 molecule into C3b and C3a fragments. The C3a fragment plays a major role in immunity by inducing chemotaxis of eosinophils and mast cells and stimulating the respiratory burst in leukocytes. Although this anaphylotoxin has been well studied in mammals, there is currently a lack of information about the structure and function of C3a anaphylotoxins in non-mammalian vertebrate species. Therefore, in the present study, we have isolated and characterized three different C3a anaphylatoxin molecules from rainbow trout, a teleost fish. C3a was generated from the trout C3 1, C3-3, and C3-4 isoforms by incubating each individual C3 molecule with purified trout factor B/C2 and factor D in the presence of Mg2+, then purifying the resulting C3a molecules by gel filtration. SDS-PAGE, N-terminal sequence and mass spectrometric analysis demonstrated the high degree of purity and expected molecular masses of the three C3a molecules. We showed that although activated trout serum was able to induce complement-dependent chemotaxis in trout head kidney leukocytes, none of the three isolated C3a molecules induced chemotaxis in the same cells. In contrast, all three C3a molecules strongly stimulated the respiratory burst of head kidney leukocytes in a dose-dependent manner. When the carboxy-terminal Arg was removed from all three C3a molecules, their ability to induce the respiratory burst was lost. These studies, therefore, provide strong evidence for the existence of three functional C3a molecules in a non-mammalian vertebrate species and suggest that some of the basic mechanisms of action of the C3a molecule have been conserved for more than 300 million years. PMID- 15043950 TI - Developing neonatal rabbit appendix, a primary lymphoid organ, is seeded by immature blood-borne B cells: evidence for roles for CD62L/PNAd, CCR7/CCL21, alpha4beta1 and LFA-1. AB - Young rabbit appendix is a homologue of chicken bursa of Fabricius; both are crucial sites for preimmune B-cell repertoire development. We describe here some of the molecules involved in the multi-step recruitment of blood-borne B cells into neonatal rabbit developing appendix. Sialyl-Lewis-x, CD62L and integrins such as LFA-1 and alpha4beta1 were detected on B cells in peripheral blood. Peripheral lymph node addressin (PNAd), a CD62L counter-receptor was observed on appendix HEV. We also detected chemokine receptor CCR7 on peripheral blood B cells and one of the CCR7 ligands, CCL21, on appendix HEV but not in appendix follicles. Higher levels of CXCR5 expression compared to CCR7 on appendix B cells suggest that CXCR5 may be involved in recruitment of B cells into follicles. The proportions of appendix B cells expressing CD62L, sialyl-Lewis-x and alpha4beta1 declined between day 3 and 4 weeks after birth while percentages of Lewis-x+ appendix B cells increased. These changes correlate with the stage of repertoire diversification by gene conversion in both rabbits and chickens. The cross reactivity of antibodies to mouse or human adhesion molecules described in this study indicates that some of the structures of these important molecules are conserved across species. PMID- 15043951 TI - The role of macrophages in the removal of apoptotic B-cells in the sheep ileal Peyer's patch. AB - In the process of generating the cells that populate the sheep's B-cell pool, the ileal Peyer's patch (PP) produces an immense number of B-cells and then destroys most of them by apoptosis. Rapid clearance of these apoptotic cells is essential for tissue homeostasis and for preventing pathology. Macrophages comprise a small percentage of cells in the follicles. They resemble macrophages found in other tissues and can be identified by the expression of MHC Class II and CD14. In this study, enriched macrophages co-cultured with apoptotic ileal PP cells showed increased DNA content as they ingested apoptotic cells. The higher the proportion of apoptotic cells in culture the greater the increase in DNA content of the macrophages. This occurred when B-cell apoptosis was initiated by a period in culture or in response to treating the animals with steroids. Thus, macrophages resident in the ileal PP follicle mediate the phagocytosis and removal of discarded B-cells. PMID- 15043952 TI - Why are French scientists striking? PMID- 15043953 TI - Growing pains in the pursuit of diabetes prevention. PMID- 15043954 TI - Cross-species transmissions of simian retroviruses in Africa and risk for human health. PMID- 15043955 TI - Purging the worm: management of Taenia solium taeniasis. PMID- 15043956 TI - Why blame systems for unsafe care? PMID- 15043957 TI - Ways around the NHS. PMID- 15043958 TI - Endarterectomy for symptomatic carotid stenosis in relation to clinical subgroups and timing of surgery. AB - BACKGROUND: Carotid endarterectomy reduces the risk of stroke in patients with recently symptomatic stenosis. Benefit depends on the degree of stenosis, and we aimed to see whether it might also depend on other clinical and angiographic characteristics, and on the timing of surgery. METHODS: We analysed pooled data from the European Carotid Surgery Trial and North American Symptomatic Carotid Endarterectomy Trial. The risk of ipsilateral ischaemic stroke for patients on medical treatment, the perioperative risk of stroke and death, and the overall benefit from surgery were determined in relation to seven predefined and seven post hoc subgroups. RESULTS: 5893 patients with 33000 patient-years of follow-up were analysed. Sex (p=0.003), age (p=0.03), and time from the last symptomatic event to randomisation (p=0.009) modified the effectiveness of surgery. Benefit from surgery was greatest in men, patients aged 75 years or older, and those randomised within 2 weeks after their last ischaemic event, and fell rapidly with increasing delay. For patients with 50% or higher stenosis, the number of patients needed to undergo surgery (ie, number needed to treat) to prevent one ipsilateral stroke in 5 years was nine for men versus 36 for women, five for age 75 years or older versus 18 for younger than 65 years, and five for those randomised within 2 weeks after their last ischaemic event, versus 125 for patients randomised after more than 12 weeks. These results were consistent across the individual trials. INTERPRETATION: Benefit from endarterectomy depends not only on the degree of carotid stenosis, but also on several other clinical characteristics such as delay to surgery after the presenting event. Ideally, the procedure should be done within 2 weeks of the patient's last symptoms. PMID- 15043960 TI - Naturally acquired simian retrovirus infections in central African hunters. AB - BACKGROUND: Hunting and butchering of wild non-human primates infected with simian immunodeficiency virus (SIV) is thought to have sparked the HIV pandemic. Although SIV and other primate retroviruses infect laboratory workers and zoo workers, zoonotic retrovirus transmission has not been documented in natural settings. We investigated zoonotic infection in individuals living in central Africa. METHODS: We obtained behavioural data, plasma samples, and peripheral blood lymphocytes from individuals living in rural villages in Cameroon. We did serological testing, PCR, and sequence analysis to obtain evidence of retrovirus infection. FINDINGS: Zoonotic infections with simian foamy virus (SFV), a retrovirus endemic in most Old World primates, were identified in people living in central African forests who reported direct contact with blood and body fluids of wild non-human primates. Ten (1%) of 1099 individuals had antibodies to SFV. Sequence analysis from these individuals revealed three geographically independent human SFV infections, each of which was acquired from a distinct non human primate lineage: De Brazza's guenon (Cercopithecus neglectus), mandrill (Mandrillus sphinx), and gorilla (Gorilla gorilla), two of which (De Brazza's guenon and mandrill) are naturally infected with SIV. INTERPRETATION: Our findings show that retroviruses are actively crossing into human populations, and demonstrate that people in central Africa are currently infected with SFV. Contact with non-human primates, such as happens during hunting and butchering, can play a part in the emergence of human retroviruses and the reduction of primate bushmeat hunting has the potential to decrease the frequency of disease emergence. PMID- 15043959 TI - European Nicotinamide Diabetes Intervention Trial (ENDIT): a randomised controlled trial of intervention before the onset of type 1 diabetes. AB - BACKGROUND: Results of studies in animals and human beings suggest that type 1 diabetes is preventable. Nicotinamide prevents autoimmune diabetes in animal models, possibly through inhibition of the DNA repair enzyme poly-ADP-ribose polymerase and prevention of beta-cell NAD depletion. We aimed to assess whether high dose nicotinamide prevents or delays clinical onset of diabetes in people with a first-degree family history of type 1 diabetes. METHOD: We did a randomised double-blind placebo-controlled trial of nicotinamide in 552 relatives with confirmed islet cell antibody (ICA) levels of 20 Juvenile Diabetes Federation (JDF) units or more, and a non-diabetic oral glucose tolerance test. Participants were recruited from 18 European countries, Canada, and the USA, and were randomly allocated oral modified release nicotinamide (1.2 g/m2) or placebo for 5 years. Random allocation was done with a pseudorandom number generator and we used size balanced blocks of four and stratified by age and national group. Primary outcome was development of diabetes, as defined by WHO criteria. Analysis was done on an intention-to-treat basis. FINDINGS: There was no difference in the development of diabetes between the treatment groups. Of 159 participants who developed diabetes in the course of the trial, 82 were taking nicotinamide and 77 were on placebo. The unadjusted hazard ratio for development of diabetes was 1.07 (95% CI 0.78-1.45; p=0.69), and the hazard ratio adjusted for age-at-entry, baseline glucose tolerance, and number of islet autoantibodies detected was 1.01 (0.73-1.38; p=0.97). Of 168 (30.4%) participants who withdrew from the trial, 83 were on placebo. The number of serious adverse events did not differ between treatment groups. Nicotinamide treatment did not affect growth in children or first-phase insulin secretion. INTERPRETATION: Large-scale controlled trials of interventions designed to prevent the onset of type 1 diabetes are feasible, but nicotinamide was ineffective at the dose we used. PMID- 15043962 TI - Salmonella typhimurium brain abscess. PMID- 15043961 TI - Interaction between heptad repeat 1 and 2 regions in spike protein of SARS associated coronavirus: implications for virus fusogenic mechanism and identification of fusion inhibitors. AB - BACKGROUND: Studies on the fusion-inhibitory peptides derived from the heptad repeat 1 and 2 (HR1 and HR2) regions of the HIV-1 envelope glycoprotein gp41 provided crucial information on the viral fusogenic mechanism. We used a similar approach to study the fusogenic mechanism of severe-acute-respiratory-syndrome associated coronavirus (SARS-CoV). METHODS: We tested the inhibitory activity against infection of two sets of peptides corresponding to sequences of SARS-CoV spike protein HR1 and HR2 regions and investigated the interactions between the HR1 and HR2 peptides by surface plasmon resonance, sedimentation equilibration analysis, circular dichroism, native polyacrylamide-gel electrophoresis, size exclusion high-performance liquid chromatography, and computer-aided homology modelling and molecule docking analysis. FINDINGS: One peptide, CP-1, derived from the HR2 region, inhibited SARS-CoV infection in the micromolar range. CP-1 bound with high affinity to a peptide from the HR1 region, NP-1. CP-1 alone had low alpha-helicity and self-associated to form a trimer in phosphate buffer (pH 7.2). CP-1 and NP-1 mixed in equimolar concentrations formed a six-helix bundle, similar to the fusogenic core structure of HIV-1 gp41. INTERPRETATION: After binding to the target cell, the transmembrane spike protein might change conformation by association between the HR1 and HR2 regions to form an oligomeric structure, leading to fusion between the viral and target-cell membranes. At the prefusion intermediate state, CP-1 could bind to the HR1 region and interfere with the conformational changes, resulting in inhibition of SARS-CoV fusion with the target cells. CP-1 might be modifiable to increase its anti-SARS-CoV activity and could be further developed as an antiviral agent for treatment or prophylaxis of SARS-CoV infection. PMID- 15043963 TI - Tongue necrosis and pericarditis. PMID- 15043964 TI - Species identification after treatment for human taeniasis. AB - Identification of species of human tapeworms is crucial because the consequences of infection by Taenia solium and T saginata are very different. However, evacuation of species-identifiable tapeworms is uncommon and Taenia spp eggs are indistinguishable under the microscope. Treatment of taeniasis consists of niclosamide followed by a purgative. Recently, we adopted preniclosamide and postniclosamide electrolyte-polyethyleneglycol salt (EPS) purges to improve bowel cleaning. Retrospective comparison of traditional castor oil with EPS purge showed that recovery of the tapeworm scolex was significantly improved (20 of 68 vs none of 46, p=0.0001) in the EPS group. Furthermore, 42 of 68 (62%) individuals receiving EPS excreted identifiable gravid proglottids. EPS treatment helps the visual identification of Taenia spp. PMID- 15043965 TI - Rabies and other lyssavirus diseases. AB - The full scale of the global burden of human rabies is unknown, owing to inadequate surveillance of this fatal disease. However, the terror of hydrophobia, a cardinal symptom of rabies encephalitis, is suffered by tens of thousands of people each year. The recent discovery of enzootic European bat lyssavirus infection in the UK is indicative of our expanding awareness of the Lyssavirus genus. The main mammalian vector species vary geographically, so the health problems created by the lyssaviruses and their management differ throughout the world. The methods by which these neurotropic viruses hijack neurophysiological mechanisms while evading immune surveillance is beginning to be unravelled by, for example, studies of molecular motor transport systems. Meanwhile, enormous challenges remain in the control of animal rabies and the provision of accessible, appropriate human prophylaxis worldwide. PMID- 15043966 TI - Challenges in the care of the acutely ill. AB - Health care providers, hospital administrators, and politicians face competing challenges to reduce clinical errors, control expenditure, increase access and throughput, and improve quality of care. The safe management of the acutely ill inpatient presents particular difficulties. In the first of five Lancet articles on this topic we discuss patients' safety in the acute hospital. We also present a framework in which responsibility for improvement and better integration of care can be considered at the level of patient, local environment, hospital, and health care system; and the other four papers in the series will examine in greater detail methods for measuring, monitoring, and improving inpatient safety. PMID- 15043967 TI - Fatigue in neurological disorders. AB - Chronic fatigue is a typical symptom of neurological diseases, and is most disabling in multiple sclerosis, postpoliomyelitis, poststroke, and in chronic fatigue syndrome. Disorders of neuromuscular junction transmission and metabolic diseases cause muscle fatigability, which is characterised by failure to sustain the force of muscle contraction (peripheral fatigue). Fatigue is also seen in diseases that affect the central, peripheral, and autonomic nervous systems (central fatigue). Enhanced perception of effort and limited endurance of sustained physical and mental activities are the main characteristics of central fatigue. Metabolic and structural lesions that disrupt the usual process of activation in pathways interconnecting the basal ganglia, thalamus, limbic system, and higher cortical centre are implicated in the pathophysiological process of central fatigue. A state of pre-existing relative hypocortisolaemia might sensitise the hypothalamic-pituitary-adrenal axis to development of persistent central fatigue after stress. The contributions of physiological, cognitive, and affective changes underlying fatigue are variable, and treatment is largely symptomatic and rehabilitative. PMID- 15043968 TI - Surprising post mortems. PMID- 15043970 TI - Securing reproductive rights. PMID- 15043969 TI - Securing reproductive rights. PMID- 15043971 TI - Securing reproductive rights. PMID- 15043972 TI - Securing reproductive rights. PMID- 15043973 TI - Securing reproductive rights. PMID- 15043974 TI - Securing reproductive rights. PMID- 15043975 TI - Erythropoietin to treat anaemia in patients with head and neck cancer. PMID- 15043976 TI - Erythropoietin to treat anaemia in patients with head and neck cancer. PMID- 15043978 TI - Erythropoietin to treat anaemia in patients with head and neck cancer. PMID- 15043979 TI - Bedsharing and sudden infant death syndrome (SIDS) in Scotland, UK. PMID- 15043980 TI - The laws of diminishing objectivity. PMID- 15043981 TI - Sleep deprivation. PMID- 15043982 TI - Junk food. PMID- 15043983 TI - Grave secrets. PMID- 15043984 TI - Methylnaltrexone prevents morphine-induced kaolin intake in the rat. AB - Opioids are frequently used analgesics, and emesis is a common opioid-induced adverse effect. Methylnaltrexone, a peripheral opioid antagonist, has the potential to block the undesired effects of opioids that are mediated by peripheral receptors while sparing the analgesic effect. We used a rat model of simulated emesis or pica to study if methylnaltrexone decreases morphine induced kaolin consumption. We observed that after morphine administration, kaolin intake increased significantly compared to intake in the vehicle group, and the increase could be attenuated by ondansetron administration. Methylnaltrexone dose dependently reduced kaolin ingestion induced by morphine. Morphine and methylnaltrexone did not significantly affect food intake and body weight in the experimental animals. Our data suggest that methylnaltrexone has therapeutic value in treating opioid-induced nausea and vomiting. PMID- 15043985 TI - Expression and functional characterization of recombinant human HDAC1 and HDAC3. AB - Histone deacetylases (HDACs) are a family of enzymes involved in transcription regulation. HDACs are known to play key roles in the regulation of cell proliferation; consequently, inhibition of HDACs has become an interesting approach for anti-cancer therapy. However, expression of mammalian HDACs has proven to be difficult. All attempts to express these HDACs in E.coli, Pichia and baculovirus systems were unsuccessful. Here we present the stable expression of human recombinant His-tagged HDAC1 and HDAC3 in mammalian cells. Full-length human genes for HDAC1 and HDAC3 were cloned into the pcDNA 3.1 vector containing a N-terminal His-tag with an enterokinase cleavage site. Recombinant HDAC enzyme activity was only detected after nickel affinity purification due to high activity of endogenous HDACs; and removal of the His-tag increased activity 2-4 fold. Western blots demonstrated the nickel affinity purified rhHDAC1 preparation also contained endogenous HDAC2 and HDAC3; likewise, rhHDAC3 preparation contained endogenous HDAC1 and HDAC2. Therefore, the active HDAC preparation is actually a multi-protein and a multi-HDAC containing complex. This provides one explanation for the similar IC50 values exhibited by SAHA and MS-275 against nuclear HDACs and rhHDAC1 and 3 preparations. These results demonstrate that recombinant forms of the HDACs can be over-expressed in mammalian cells, isolated as active multi-protein complexes that contain multiple HDAC enzymes, and caution must be used when determining HDAC inhibitor in vitro selectivity. PMID- 15043986 TI - Trans fatty acids in hydrogenated fat inhibited the synthesis of the polyunsaturated fatty acids in the phospholipid of arterial cells. AB - Our hypothesis that the trans fatty acids in hydrogenated fat inhibited the synthesis of polyunsaturated fatty acids in the phospholipid of arterial cells was tested with five groups each with six pregnant porcine fed from d 35 of gestation and during lactation. The basal diet contained 2% corn oil (control). The other four diets included the control + 10% butter or 10% hydrogenated fat plus two levels of Mg. Plasma, milk and aortic phospholipid fatty acids, phospholipid composition and calcium content of the aorta from the piglets were determined. At 48 +/- 2 d of age, the aorta phospholipid of piglets from porcine fed hydrogenated fat contained a significantly higher concentration of linoleic acid, less arachidonic acid, and less long chain polyunsaturated fatty acid (PUFA) than did piglets from porcine fed either butterfat or the control diet. Mg had no effect. These changes in composition in piglets from porcine fed hydrogenated fat indicate that trans fat inhibits the metabolic conversion of linoleic acid to arachidonic acid and to other n-6 PUFA. The aortic calcium content data showed a significant interaction of calcium concentration with age. We concluded: 1) that dietary trans fat perturbed essential fatty acid (EFA) metabolism which led to changes in the phospholipid fatty acid composition in the aorta, the target tissue of atherogenesis, 2) this inhibition of EFA to PUFA by the isomeric fatty acids in hydrogenated fat is a risk factor in the development of coronary heart disease. PMID- 15043987 TI - Intermittent nicotine administration modulates food intake in rats by acting on nicotine receptors localized to the brainstem. AB - Previous studies have shown nicotine (NIC) administration leads to decreased food intake, while other investigations have reported that NIC stimulates c-Fos expression in the brainstem. Whether there is a causal relationship between NIC effects on ingestion and its effect on brainstem neurons is uncertain, however we hypothesized that blocking NIC action in the brainstem would prevent, to some extent, the hypophagic effects of NIC. In the present study, cannulas were placed in the fourth ventricle of rats. A dose of NIC or saline was injected i.p. in four equal injections during the dark phase for four days. At the start of the second day of injections the NIC receptor antagonist mecamylamine (MEC) or artificial cerebrospinal fluid (a-CSF) was infused intracerebroventricularly (i.c.v.). Thus, four experimental groups were examined: a-CSF + SAL; a-CSF + NIC; MEC + SAL; MEC + NIC. Meal patterns were recorded using a computerized system and water intake and body weight were measured daily. Peripheral NIC injections suppressed food intake by decreasing meal size, whereas infusion of the NIC receptor antagonist MEC (4 microg) into the fourth ventricle blocked the NIC suppression of food intake. Moreover, the MEC effect was due primarily to an increase in dark phase meal size, which suggests neurons localized to the brainstem transmit NIC signals that regulate feeding behavior by affecting meal size. PMID- 15043988 TI - Effects of the dietary supplements, activated charcoal and copper chlorophyllin, on urinary excretion of trimethylamine in Japanese trimethylaminuria patients. AB - Trimethylaminuria (TMAU) is a metabolic disorder characterized by the inability to oxidize and convert dietary-derived trimethylamine (TMA) to trimethylamine N oxide (TMAO). This disorder has been relatively well-documented in European and North American populations, but no reports have appeared regarding patients in Japan. We identified seven Japanese individuals that showed a low metabolic capacity to convert TMA to its odorless metabolite, TMAO. The metabolic capacity, as defined by the concentration of TMAO excreted in the urine divided by TMA concentration plus TMAO concentration, in these seven individuals ranged from 70 to 90%. In contrast, there were no healthy controls examined with less than 95% of the metabolic capacity to convert TMA to TMAO. The intake of dietary charcoal (total 1.5 g charcoal per day for 10 days) reduced the urinary free TMA concentration and increased the concentration of TMAO to normal values during charcoal administration. Copper chlorophyllin (total 180 mg per day for 3 weeks) was also effective at reducing free urinary TMA concentration and increasing TMAO to those of concentrations present in normal individuals. In the TMAU subjects examined, the effects of copper chlorophyllin appeared to last longer (i.e., several weeks) than those observed for activated charcoal. The results suggest that the daily intake of charcoal and/or copper chlorophyllin may be of significant use in improving the quality of life of individuals suffering from TMAU. PMID- 15043989 TI - A protective effect of the synthetic coumarine derivative Cloricromene against DNB-colitis in the rat. AB - Biologic therapies, namely antibodies against tumor necrosis factor-alpha (TNF- alpha) or its receptors, have been recently introduced for the treatment of patients with inflammatory bowel disease (IBD). In the present study the effects of cloricromene, an agent with known antithrombotic actions and with demonstrated anti-TNF- alpha activity were investigated in a rat model of experimental colitis induced with dinitrobenzenesulphonic acid (DNB)/ethanol. We investigated three experimental groups: (i) sham-colitis with vehicle-treatment (controls, n = 6), (ii) colitis with vehicle-treatment (saline, 0.1 ml s.c., daily) (DNB-V, n = 7), (iii) colitis with cloricromene-treatment (10 mg/kg/day s.c.; DNB-C, n = 8). After 7 days, the weight gain, colon wet weight, macroscopic damage score, coagulation parameters, colon mucosal myeloperoxidase activity (MPO), and tissue concentrations of TNF- alpha and of macrophage inhibitory peptide-2 (MIP-2) were assessed. The macroscopic damage scores, colon wet weights, and tissue MIP-2 levels were significantly increased in untreated and in cloricromene-treated rats compared with controls. Cloricromene treatment was associated with a minor body weight loss (p < 0.025) and significantly reduced tissue concentrations of MPO and TNF-alpha (p < 0.02, both). Blood coagulation parameters were not affected by treatment. In the DNB-model treatment with cloricromene effectively reduces tissue levels of TNF- alpha and of myeloperoxidase, whereas MIP-2 concentrations were not influenced. Blood coagulation parameters remained unchanged indicating safety of treatment. Since biological therapies frequently fail to improve disease course of IBD, other therapies with similar targets should be further investigated. PMID- 15043990 TI - Chronic nitric oxide exposure alters the balance between endothelium-derived relaxing factors released from rat renal arteries: prevention by treatment with NOX-100, a NO scavenger. AB - We investigated whether nitric oxide (NO) exposure alters the balance between NO and endothelium-derived hyperpolarizing factor (EDHF) released from rat renal arteries. To produce states of acutely or chronically excessive NO, lipopolysaccharide (LPS) was administered intraperitoneally to rats in a single dose of 4 mg/kg (LPS-single group) or in stepwise doses of 0.5, 1.0 and 2.0 mg/kg every other day (LPS-repeated group). On the day after LPS treatment, the protein levels of inducible NO synthase (iNOS) and endothelial NOS (eNOS) were measured, and the relaxation responses were determined in the renal arteries. The protein levels of iNOS markedly increased in both LPS-treated groups, while those of eNOS significantly increased in the LPS-repeated group compared with those in the respective control groups. In both LPS-treated groups, the relaxations in response to acetylcholine (ACh) and sodium nitroprusside remained unchanged. The ACh-induced relaxations in the presence of N(G)-nitro-L-arginine methyl ester, a NOS inhibitor, or by 1H-[1, 2, 4-] oxadiazole [4, 3-a] quinoxalin-1-one, a soluble guanylyl cyclase inhibitor, i.e. EDHF-mediated relaxations were significantly impaired in the LPS-repeated group but not in the LPS-single group, indicating increase in NO-mediated relaxation in the LPS-repeated group. These changes in the protein levels and EDHF-mediated relaxations induced by ACh observed in the LPS-repeated group were restored by treatment with NOX-100, a NO scavenger. These results suggest that persistent but not acute excessive NO exposure in rats impairs EDHF-mediated relaxation in renal arteries, leading to a compensatory upregulation of the eNOS/NO pathway. PMID- 15043991 TI - Inhibition of cytochrome P450 activities by oleanolic acid and ursolic acid in human liver microsomes. AB - Oleanolic acid (OA) and ursolic acid (UA), triterpene acids having numerous pharmacological activities including anti-inflammatory, anti-cancer, and hepato protective effects, were tested for their ability to modulate the activities of several cytochrome P450 (CYP) enzymes using human liver microsomes. OA competitively inhibited CYP1A2-catalyzed phenacetin O-deethylation and CYP3A4 catalyzed midazolam 1-hydroxylation, the major human drug metabolizing CYPs, with IC50 (Ki) values of 143.5 (74.2) microM and 78.9 (41.0) microM, respectively. UA competitively inhibited CYP2C19-catalyzed S-mephenytoin 4'-hydroxylation with an IC50 (Ki) value of 119.7 (80.3) microM. However, other CYPs tested showed no or weak inhibition by both OA and UA. The present study demonstrates that OA and UA have inhibitory effects on CYP isoforms using human liver microsomes. It is thus likely that consumption of herbal medicines containing OA or UA, or administration of OA or UA, can cause drug interactions in humans when used concomitantly with drugs that are metabolized primarily by CYP isoforms. In addition, it appears that the inhibitory effect of OA on CYP1A2 is, in part, related to its anti-inflammatory and anticancer activities. PMID- 15043992 TI - Secoisolariciresinol and isotaxiresinol inhibit tumor necrosis factor-alpha dependent hepatic apoptosis in mice. AB - The effects of secoisolariciresinol (1) and isotaxiresinol (2), two major lignans isolated from the wood of Taxus yunnanensis, on tumor necrosis factor-alpha (TNF alpha)-dependent hepatic apoptosis induced by D-galactosamine (d GalN)/lipopolysaccharide (LPS) were investigated in mice. Co-administration of d GalN (700 mg/kg) and LPS (10 microg/kg) resulted in a typical hepatic apoptosis characterized by DNA fragmentation and the formation of apoptotic bodies. Serum glutamic pyruvic transaminase (sGPT) and glutamic oxaloacetic transaminase (sGOT) levels were also raised at 8 h after d-GalN/LPS intoxication due to a severe necrosis of hepatocytes. Pre-administration of 1 or 2 (50, 10 mg/kg, i.p.) 12 and 1 h before d-GalN/LPS significantly reduced DNA fragmentation and prevented chromatin condensation, apoptotic body formation and hepatitis. Pro-inflammatory cytokines such as TNF-alpha and interferon-gamma (IFN-gamma) secreted from LPS activated macrophages are important mediators of hepatocyte apoptosis in this model. Pre-treatment with 1 or 2 significantly inhibited the elevation of serum TNF-alpha and IFN-gamma levels. In a separate experiment, both lignans had a significant dose-dependent protective effect on d-GalN/TNF-alpha-induced cell death in primary cultured mouse hepatocytes and TNF-alpha-mediated cell death in murine L929 fibrosarcoma cells. These results indicated that 1 and 2 prevent d GalN/LPS-induced hepatic injury by inhibiting hepatocyte apoptosis through the blocking of TNF-alpha and IFN-gamma production by activated macrophages and direct inhibition of the apoptosis induced by TNF-alpha. PMID- 15043993 TI - Regulation of calcium signalling by 1-O-alkylglycerols in human Jurkat T lymphocytes. AB - We studied the role of natural occurring 1-O-alkylglycerols on the calcium signalling in Jurkat T-cells. Alkylglycerols evoked an increase in free intracellular calcium concentration [Ca2+]i, in a dose-dependent manner. When the experiments were performed in calcium-free buffer, the alkylglycerol response on the rise of [Ca2+]i was wholly abolished compared with the one in calcium containing buffer, suggesting that these etherlipids induce a calcium influx by the opening of Ca2+ channels. We further employed inhibitors of voltage-gated calcium channels. We observed that omega-conotoxin, a blocker of N-type voltage activated Ca2+ channels, but not verapamil, a blocker of L-type voltage-activated Ca2+ channels, curtailed significantly the calcium rise evoked by the lipid agents. Alkylglycerols also induced plasma membrane depolarisation, known to be involved in the opening of the voltage-gated calcium channels. Our study shows that alkylglycerols increase [Ca2+]i influx in human Jurkat T-cells possibly by modulating the permeability of calcium channels. PMID- 15043994 TI - Dynactins p25 and p27 are predicted to adopt the LbetaH fold. AB - Dynactin is a multimeric protein essential for the minus-end-directed transport driven by microtubule-based motor dynein. The pointed-end subcomplex in dynactin contains p62, p27, p25, and Arp11 subunits, and is thought to participate in interactions with membranous cargoes. We used sequence and structure prediction analysis to study dynactins p25 and p27. Here we present evidence that strongly supports that dynactins p27 and p25 contain the isoleucine-patch motif and adopt the left-handed parallel beta-helix fold. The structural models we obtained could contribute to the understanding of the complex interactions that dynactins are able to establish with cargo particles, microtubules or other dynactin subunits. PMID- 15043995 TI - Anti-amyloid activity of neprilysin in plaque-bearing mouse models of Alzheimer's disease. AB - Abnormally high concentrations of beta-amyloid peptide (Abeta) and amyloid plaque formation in Alzheimer's disease (AD) may be caused either by increased generation or by decreased degradation of Abeta. Therefore, activation of mechanisms that lower brain Abeta levels is considered valuable for AD therapy. Neuronal upregulation of neprilysin (NEP) in young transgenic mice expressing the AD-causing amyloid precursor protein mutations (SwAPP) led to reduction of brain Abeta levels and delayed Abeta plaque deposition. In contrast, a comparable increase of brain NEP levels in aged SwAPP mice with pre-existing plaque pathology did not result in a significant reduction of plaque pathology. Therefore, we suggest that the potential of NEP for AD therapy is age-dependent and most effective early in the course of AD pathophysiology. PMID- 15043996 TI - A thermostable manganese-containing superoxide dismutase from pathogen Chlamydia pneumoniae. AB - The gene CP0718 encoding a putative manganese-containing superoxide dismutase of Chlamydia pneumoniae AR39 was cloned and expressed in Escherichia coli. Characterization showed that the expressed protein with a monomeric molecular mass of 23.1 kDa had superoxide dismutase (SOD) activity and the cofactor of CpSOD was a bivalent manganese cation. It is unexpected that this enzyme was hyperthermostable, and maintained about 90% activity after incubation at 70 degrees C for 60 min. Manganese binding residues found in the SOD sequences from different species are conserved in CpSOD. Bioinformatics analysis compared with Propionibacterium shermanii MnSOD was performed to elucidate the CpSOD hyperthermostability based on amino acid sequences. PMID- 15043997 TI - Characterization of the human STAT5A and STAT5B promoters: evidence of a positive and negative mechanism of transcriptional regulation. AB - We recently published the genomic characterization of the STAT5A and STAT5B paralogous genes that are located head to head in the 17q21 chromosome and share large regions of sequence identity. We here demonstrate by transient in vitro transfection that STAT5A and STAT5B promoters are able to direct comparable levels of transcription. The expression of basal promoters is enhanced after Sp1 up-regulation in HeLa and SL2 cells while DNA methylation associated to the recruitment of MeCP2 methyl CpG binding protein down-regulates STAT5A and B promoters by interfering with Sp1-induced transcription. In addition, cross species sequence comparison identified a bi-directional negative cis-acting regulatory element located in the STAT5 intergenic region. PMID- 15043998 TI - The nitration of tau protein in neurone-like PC12 cells. AB - Tyrosine nitration of proteins is emerging as a post-translational modification playing a role in physiological conditions. Looking for the molecular events triggered by nitric oxide in nerve growth factor-induced neuronal differentiation, we now find that nitration occurs on the microtubule-associated protein tau. In differentiated PC12 cells, we have identified as tau a nitrated protein that co-immunoprecipitates with alpha-tubulin and indicated that the modified protein is associated with the cytoskeleton but it is confined to a restricted cell region. This paper supplies the first evidence that nitration of tau occurs in a physiological process and suggests that it could play a role in neuronal differentiation. PMID- 15043999 TI - Degadration of mismatch repair hMutSalpha heterodimer by the ubiquitin-proteasome pathway. AB - Mismatch repair plays a critical role in genome stability. This process requires several proteins including hMSH2/hMSH6 (hMutSalpha) heterodimer involved in the first stage of the process, the mispair recognition. We previously reported that in U937 and HL-60 cell lines, hMSH2 and hMSH6 protein expression was much lower than that in HeLa and KG1a cells. Here, we showed that the decreased expression of hMutSalpha results from differences in the degradation rate of both proteins by the ubiquitin-proteasome pathway. Our data suggest that in human cell lines, ubiquitin-proteasome could play an important role in the regulation of hMutSalpha protein expression, thereby regulating mismatch repair activity. PMID- 15044000 TI - Applied jasmonates accumulate extracellularly in tomato, but intracellularly in barley. AB - Jasmonic acid (JA) and its derivatives are well-characterized signaling molecules in plant defense and development, but the site of their localization within plant tissue is entirely unknown. To address the question whether applied JA accumulates extracellularly or intracellularly, leaves of tomato and barley were fed with 14C-labeled JA and the label was localized in cryofixed and lyophilized leaf tissues by microautoradiography. In tomato the radioactivity was detectable within the apoplast, but no label was found within the mesophyll cells. By contrast, in barley leaf tissues, radioactivity was detected within the mesophyll cells suggesting a cellular uptake of exogenously applied JA. JA, applied to leaves of both plants as in the labeling experiments, led in all leaf cells to the expression of JA-inducible genes indicating that the perception is completed by JA signal transduction. PMID- 15044001 TI - Polyvalency of Tn (GalNAcalpha1-->Ser/Thr) glycotope as a critical factor for Vicia villosa B4 and glycoprotein interactions. AB - Vicia villosa B(4) (VVL-B(4)) is an important lectin for detecting exposed Tn (GalNAcalpha1-Ser/Thr) determinants on cancer cells. In order to elucidate the binding factors involved in VVL-B(4) and glycotope interaction, the binding properties of this lectin were analyzed by enzyme-linked lectinosorbent and inhibition assays. From the results, it is concluded that the most critical factor affecting VVL-B(4) binding is polyvalency at the alpha anomer of Gal with NH CH(3)CO at carbon-2 (Tn epitope), which enhances the reactivity by 3.3x10(5) times over monovalent Gal. The reactivities of glycotopes can be ranked as follows: high density Tn cluster >>Tn glycopeptides (MW<3.0x10(3) >> monomeric Tn to tri- Tn glycopeptides >>> other GalNAcalpha/beta-related structural units>Gal and Galalpha- or beta-linked ligands, demonstrating the essential role of the polyvalency of Tn glycotopes in the enhancement of the binding. PMID- 15044002 TI - Important role of Ser443 in different thermal stability of human glutamate dehydrogenase isozymes. AB - Molecular biological studies confirmed that two glutamate dehydrogenase isozymes (hGDH1 and hGDH2) of distinct genetic origin are expressed in human tissues. hGDH1 is heat-stable and expressed widely, whereas hGDH2 is heat-labile and specific for neural and testicular tissues. A selective deficiency of hGDH2 has been reported in patients with spinocerebellar ataxia. We have identified an amino acid residue involved in the different thermal stability of human GDH isozymes. At 45 degrees C (pH 7.0), heat inactivation proceeded faster for hGDH2 (half life=45 min) than for hGDH1 (half-life=310 min) in the absence of allosteric regulators. Both hGDH1 and hGDH2, however, showed much slower heat inactivation processes in the presence of 1 mM ADP or 3 mM L-Leu. Virtually most of the enzyme activity remained up to 100 min at 45 degrees C after treatment with ADP and L-Leu in combination. In contrast to ADP and L-Leu, the thermal stabilities of the hGDH isozymes were not affected by addition of substrates or coenzymes. In human GDH isozymes, the 443 site is Arg in hGDH1 and Ser in hGDH2. Replacement of Ser by Arg at the 443 site by cassette mutagenesis abolished the heat lability of hGDH2 with a similar half-life of hGDH1. The mutagenesis at several other sites (L415M, A456G, and H470R) having differences in amino acid sequence between the two GDH isozymes did not show any change in the thermal stability. These results suggest that the Ser443 residue plays an important role in the different thermal stability of human GDH isozymes. PMID- 15044003 TI - Virus survival of RNA silencing without deploying protein-mediated suppression in Nicotiana benthamiana. AB - RNA silencing represents an evolutionarily conserved defence mechanism that plays a key antiviral role in protecting plants and animals against virus infection. To counterattack, plant, animal and fungal viruses produce proteins capable of suppressing RNA silencing. Here, we report an unprecedented phenomenon that Potato virus X, a single-stranded positive RNA virus, is able to survive RNA silencing without deploying protein-mediated anti-silencing by revealing an unexpected symptom re-emergence and re-accumulation of viral RNAs and proteins in plants maintaining strong RNA silencing. Our results provide evidence that a plant virus may have developed a getaway strategy to survive RNA silencing. PMID- 15044004 TI - LRP-1/TbetaR-V mediates TGF-beta1-induced growth inhibition in CHO cells. AB - The type V transforming growth factor-beta (TGF-beta) receptor (TbetaR-V) is hypothesized to be involved in cellular growth inhibition by TGF-beta(1). Recently, TbetaR-V was found to be identical to low density lipoprotein receptor related protein-1 (LRP-1). Here we demonstrate that TGF-beta(1) inhibits growth of wild-type CHO cells but not LRP-1-deficient mutant cells (CHO-LRP-1(-) cells). Stable transfection of CHO-LRP-1(-) cells with LRP-1 cDNA restores the wild-type morphology and the sensitivity to growth inhibition by TGF-beta(1). In addition, overexpression of LRP-1 minireceptors exerts a dominant negative effect and attenuates the growth inhibitory response to TGF-beta(1) in wild-type CHO cells. These results suggest that LRP-1/TbetaR-V is critical for TGF-beta(1)-mediated growth inhibition in CHO cells. PMID- 15044005 TI - Identification of a novel class of annexin genes. AB - The annexins are a family of calcium- and phospholipid-binding proteins that have been widely studied in animals. Investigation of annexins in the fungus Aspergillus fumigatus identified a novel annexin-like gene (ANXC4) as well as two conventional annexins (ANXC3.1 and ANXC3.2). The genes were initially identified by bioinformatics, and sequences were then determined experimentally. Reverse transcription polymerase chain reaction indicated that all three genes were expressed. ANXC4 lacked calcium-binding consensus sequences and had a 553 residue N-terminal tail. However, bioinformatics indicated that ANXC4 is an annexin and homologues were identified in other filamentous fungi. ANXC4 therefore represents a new grouping within the annexin family. PMID- 15044006 TI - Killing of target cells by redirected granzyme B in the absence of perforin. AB - Granzyme B (GzmB) is a potent apoptosis-inducing serine protease of cytotoxic lymphocytes. Following receptor-mediated endocytosis, GzmB is supposed to enter the cytosol through perforin-mediated membrane disruption. We investigated whether retargeting of GzmB to Lewis Y positive surface receptors could lead to perforin-independent target cell death. We coupled recombinant GzmB to the Lewis Y-binding antibody dsFv-B3. Targeting of GzmB to Lewis Y positive cells triggered cell death with similar efficacy as dsFv-B3 targeted Pseudomonas exotoxin fragment 38 (PE38). Since GzmB was only weakly inhibited by plasma proteins, GzmB based immunoconjugates should be useful as a new class of immunotoxins with low immunogenicity utilizing programmed cell death for therapeutic purposes. PMID- 15044007 TI - Effect of N-acetylglucosaminyltransferase V on the expressions of other glycosyltransferases. AB - Transfection of sense cDNA of N-acetylglucosaminyltransferase V (GnTV) into H7721 human hepatocellular carcinoma cells resulted in the decreased expression of surface sialyl Lewis X (SLe(x)), a sialylated fucose-containing antigen. The enzymatic mechanisms were speculated to be the concomitantly decreased expression of alpha1,3-fucosyltransferase (FucT)-III, -VI, -VII and the branching enzyme of O-glycans, core 2-beta1,6-N-acetylglucosaminyltransferase (C2GnT)-I, -II. These two glycosyltransferase families were suggested to be the key enzymes in the synthesis of SLe(x). The expression of alpha2,3-sialyltransferase (ST3)-IV, but not ST3-I, -II and -III was elevated by sense GnTV. However, it did not cause the increase of SLe(x) synthesis. Transfection of antisense GnTV into H7721 cells showed entirely opposite effects on the expression of above-mentioned SLe(x) and glycosyltransferases as the sense GnTV. PMID- 15044008 TI - Cholesterol diet-induced hyperlipidemia influences gene expression pattern of rat hearts: a DNA microarray study. AB - To profile gene expression patterns involved in the direct myocardial effect of cholesterol-enriched diet-induced hyperlipidemia, we monitored global gene expression changes by DNA microarray analysis of 3200 genes in rat hearts. Twenty six genes exhibited significant up-regulation and 25 showed down-regulation in hearts of rats fed a 2% cholesterol-enriched diet for 8 weeks as compared to age matched controls. The expression changes of 12 selected genes were also assessed by real-time quantitative polymerase chain reaction. Genes with altered expression in the heart due to hyperlipidemia included procollagen type III, cofilin/destrin, tensin, transcription repressor p66, synaptic vesicle protein 2B, Hsp86, chaperonin subunit 5epsilon, metallothionein, glutathione S transferase, protein kinase C inhibitor, ATP synthase subunit c, creatine kinase, chloride intracellular channel 4, NADH oxidoreductase and dehydrogenase, fibronectin receptor beta chain, CD81 antigen, farnesyltransferase, calreticulin, disintegrin, p120 catenin, Smad7, etc. Although some of these genes have been suspected to be related to cardiovascular diseases, none of the genes has been previously shown to be involved in the mechanism of the cardiac effect of hyperlipidemia. PMID- 15044009 TI - Immediate response of the DnaK molecular chaperone system to heat shock. AB - The familiar heat shock response in cells comprises the enhanced expression of molecular chaperones. In recent experiments with the Hsp70 system of Escherichia coli, the co-chaperone GrpE has been found to undergo a reversible thermal transition in the physiological temperature range. Here, we tested whether this thermal transition is of functional significance in the complete DnaK/DnaJ/GrpE chaperone system. We found that a mere increase in temperature resulted in a higher fraction of fluorescence-labeled peptides being sequestered by DnaK. This direct adaptation of the DnaK/DnaJ/GrpE chaperone system to heat shock conditions may serve to bridge the time lag of enhanced chaperone expression. PMID- 15044010 TI - N-linked glycosylation of platelet P2Y12 ADP receptor is essential for signal transduction but not for ligand binding or cell surface expression. AB - P(2)Y(12) receptor is a G(i)-coupled adenosine diphosphate (ADP) receptor with a critical role in platelet aggregation. It contains two potential N-linked glycosylation sites at its extra cellular amino-terminus, which may modulate its activity. Studies of both tunicamycin treatment and site-directed mutagenesis have revealed a dispensable role of the N-linked glycosylation in the receptor's surface expression and ligand binding activity. However, the non-glycosylated P(2)Y(12) receptor is defective in the P(2)Y(12)-mediated inhibition of the adenylyl cyclase activity. Thus the study uncovers an unexpected vital role of N linked glycans in receptor's signal transducing step but not in surface expression or ligand binding. PMID- 15044011 TI - Gaining insight into the role of serine 282 in B. napus FAE1 condensing enzyme. AB - To gain some insight whether there is an absolute requirement for the serine 282 to yield a functional fatty acid elongase 1 condensing enzyme we have introduced point mutations in the FAE1 coding sequence which led to the substitution of serine 282 with several aliphatic or aromatic amino acids. The mutated FAE1 polypeptides were expressed in yeast. Gas chromatography analyses of the fatty acid methyl esters from yeast lysates and fatty acid elongase activity assays demonstrated that there is not an absolute requirement for serine at position 282 to yield a functional FAE1 condensing enzyme. PMID- 15044012 TI - Screening of functional antidotes of RNA aptamers against bovine thrombin. AB - A specific RNA aptamer (T705) against bovine thrombin had been obtained after seven rounds of SELEX (systematic evolution of ligands by exponential enrichment) selection from a random RNA library previously. In order to further investigate the relationship between the structure and function of this aptamer, three truncated RNA aptamers, T705a, T705b and T705c, were designed according to the secondary structure of T705 RNA. Our results showed that T705c keeping the precise stem-loop structure but lacking most of the stem region sequence of T705 could inhibit clot formation in vitro in the same way as its parental form. We also report here that single-stranded DNA (ssDNA) antisense oligonucleotides, c' and c'-22, which were complementary to different portions of T705c could act as efficient antidotes reversing the inhibitory activity of T705. It is demonstrated for the first time that ssDNA antisense oligonucleotides are potential antidotes of RNA aptamers and this may be an effective, rapid strategy to find antidotes of RNA aptamers which would be of important usefulness in basic research and drug screening. PMID- 15044013 TI - Peroxynitrite formation from the simultaneous reduction of nitrite and oxygen by xanthine oxidase. AB - One electron reductions of oxygen and nitrite by xanthine oxidase form peroxynitrite. The nitrite and oxygen reducing activities of xanthine oxidase are regulated by oxygen with K(oxygen) 26 and 100 microM and K(nitrite) 1.0 and 1.1 mM with xanthine and NADH as donor substrates. Optimal peroxynitrite formation occurs at 70 microM oxygen with purine substrates. Kinetic parameters: V(max) approximately 50 nmol/min/mg and K(m) of 22, 36 and 70 microM for hypoxanthine, pterin and nitrite respectively. Peroxynitrite generation is inhibited by allopurinol, superoxide dismutase and diphenylene iodonium. A role for this enzyme activity can be found in the antibacterial activity of milk and circulating xanthine oxidase activity. PMID- 15044014 TI - Molecular cloning and characterization of beta1,4-N acetylgalactosaminyltransferases IV synthesizing N,N'-diacetyllactosediamine. AB - A sequence highly homologous to beta1,4-N-acetylgalactosaminyltransferase III (beta4GalNAc-T3) was found in a database of human expressed sequence tags. The full-length open reading frame of the gene, beta4GalNAc-T4 (GenBank accession number AB089939), was cloned using the 5' rapid amplification of cDNA ends method. It encodes a typical type II transmembrane protein of 1039 amino acids having 42.6% identity with beta4GalNAc-T3. The recombinant enzyme transferred N acetylgalactosamine to N-acetylglucosamine-beta-benzyl with a beta1,4-linkage to form N,N'-diacetyllactosediamine as did beta4GalNAc-T3. In specificity toward oligosaccharide acceptor substrates, it was quite similar to beta4GalNAc-T3 in vitro, however, the tissue distributions of the two enzymes were quite different. These results indicated that the two enzymes have similar roles in different tissues. PMID- 15044016 TI - Analysis of GT-3a identifies a distinct subgroup of trihelix DNA-binding transcription factors in Arabidopsis. AB - Trihelix DNA-binding factors (or GT factors) bind to GT elements found in the promoters of many plant genes. Although the binding specificity and the transcriptional activity of some members (e.g. GT-1 and GT-2) have been studied, the regulatory function of this family of transcription factors remains largely unknown. In this work, we have characterised a new GT factor, namely GT-3a, and a closely related member, GT-3b. We show that (1) they can form either homo- or heterodimers but do not interact with GT-1; (2) they are predominantly expressed in floral buds and roots; (3) GT-3a cannot bind to the binding sites of GT-1 or GT-2, but binds to the cab2 and rbcS-1A gene promoters via the 5'-GTTAC sequence, which has been previously shown to be the core of the Site 1 type of GT elements. These results suggest that GT-3a and GT-3b belong to a distinct subgroup of GT factors and that each subgroup of GT factors binds to a functionally distinct type of cis-acting GT elements. PMID- 15044015 TI - Ablation of SM22alpha decreases contractility and actin contents of mouse vascular smooth muscle. AB - The actin-binding protein SM22alpha marks contractile differentiation in smooth muscle, but its function is unknown. We tested its role in arterial contractility and stretch-sensitive vascular protein synthesis. Active stress in depolarised mesenteric resistance arteries and portal veins was reduced by 40% in SM22alpha( /-) mice. Passive and active arterial circumference-force relationships were shifted leftwards, whereas alpha(1)-adrenergic responses were increased. Actin contents were 10-25% lower in vessels from SM22alpha(-/-) mice, but protein composition was otherwise similar. Synthesis of SM22alpha, calponin and alpha actin, but not beta-actin, was sensitive to stretch. Ablation of SM22alpha did not affect stretch sensitivity of any of these proteins. Thus, SM22alpha plays a role in contractility, possibly by affecting actin filament organisation. PMID- 15044017 TI - Formation of circular polyribosomes in wheat germ cell-free protein synthesis system. AB - We report a morphological study of functioning ribosomes in a efficient and robust cell-free protein synthesis system prepared from wheat embryos. Sucrose density gradient analysis of translated mixtures programmed with luciferase mRNAs having different 5' and 3' untranslated regions showed formation of large polysomes. Electron microscopic examination of translation mixtures programmed with those of capped and polyadenylated mRNA revealed that ribosomes assemble into a circular-type polysome in vitro. Furthermore, a series of experiments using mRNAs lacking either cap, poly(A) tail or both also resulted in the formation of circular polysomes, which are indistinguishable from those with the original mRNA. The wheat germ cell-free system may provide a good experimental system for understanding functional ribosomes at the molecular level. PMID- 15044018 TI - The islet-specific glucose-6-phosphatase-related protein, implicated in diabetes, is a glycoprotein embedded in the endoplasmic reticulum membrane. AB - The islet-specific glucose-6-phosphatase-related protein (IGRP) has no known catalytic activity, but is of interest because it is the source of the peptide autoantigen targeted by a prevalent population of pathogenic CD8(+) T cells in non-obese diabetic mice. To better understand the potential roles of this protein in diabetes mellitus, we examine the subcellular localization and membrane topography of human IGRP. We show that IGRP is a glycoprotein, held in the endoplasmic reticulum by nine transmembrane domains, which is degraded in cells predominantly through the proteasome pathway that generates the major histocompatibility complex class I-presented peptides. PMID- 15044019 TI - HBZ interacts with JunD and stimulates its transcriptional activity. AB - Human T-cell leukemia virus type I (HTLV-I) bZIP factor (HBZ) is a viral basic leucine zipper protein that was originally described as a partner of cAMP response element binding protein-2 and as a repressor of HTLV-I viral transcription. In addition, HBZ is able to interact with the activator protein-1 (AP-1) transcription factors c-Jun and JunB, the interaction with c-Jun leading to a transcriptional repression of AP-1-regulated genes. Here we show that HBZ also interacts with JunD in vitro and in vivo, and that this association occurs via the bZIP domain of the two proteins. Moreover, we show that HBZ can activate JunD-dependent transcription and that its amino-terminus is required. PMID- 15044020 TI - The first plant type III polyketide synthase that catalyzes formation of aromatic heptaketide. AB - A cDNA encoding a novel plant type III polyketide synthase (PKS) was cloned from rhubarb (Rheum palmatum). A recombinant enzyme expressed in Escherichia coli accepted acetyl-CoA as a starter, carried out six successive condensations with malonyl-CoA and subsequent cyclization to yield an aromatic heptaketide, aloesone. The enzyme shares 60% amino acid sequence identity with chalcone synthases (CHSs), and maintains almost identical CoA binding site and catalytic residues conserved in the CHS superfamily enzymes. Further, homology modeling predicted that the 43-kDa protein has the same overall fold as CHS. This provides new insights into the catalytic functions of type III PKSs, and suggests further involvement in the biosynthesis of plant polyketides. PMID- 15044021 TI - Susceptibility to transglutaminase of gliadin peptides predicted by a mass spectrometry-based assay. AB - A peptidomics approach was developed to identify transglutaminase-susceptible Q residues within a pepsin-trypsin gliadin digest. Based on tagging with a monodansylcadaverine fluorescent probe, six alpha/beta-, gamma-gliadin, and low molecular weight glutenin peptides were identified by nanospray tandem mass spectrometry. In functioning as an acyl acceptor, tissue transglutaminase was able to form complexes with the glutamine-rich gliadin peptides, whereas by lowering pH, the peptides were deamidated by transglutaminase at the same Q residues, which were previously transamidated. The main common feature shared by the peptides was the consensus sequence Q-X-P. Our findings offer relevant information for the understanding of how dietary peptides interact with the host organism in celiac disease. PMID- 15044023 TI - Effect of salt and osmotic stress upon expression of the ethylene receptor ETR1 in Arabidopsis thaliana. AB - In hormone perception, varying the concentrations of hormone, receptor, or downstream signaling elements can modulate signal transduction. Previous research has demonstrated that ethylene biosynthesis in plants is regulated by abiotic factors. Here we report that exposure of Arabidopsis plants to NaCl reduced expression of the ethylene receptor ETR1. The change in gene expression was reflected at the protein level based on immunoblot analysis. Further analysis supports a general effect of osmotic stress upon the expression level of ETR1. The reduction in ETR1 levels should cause increased sensitivity of the plant to ethylene. These results suggest that plant responses to abiotic stress are modulated by changes in the expression level of ethylene receptors. PMID- 15044022 TI - The crystal structure of the Escherichia coli lipocalin Blc suggests a possible role in phospholipid binding. AB - Lipocalins form a large multifunctional family of small proteins (15-25 kDa) first discovered in eukaryotes. More recently, several types of bacterial lipocalins have been reported, among which Blc from Escherichia coli is an outer membrane lipoprotein. As part of our structural genomics effort on proteins from E. coli, we have expressed, crystallized and solved the structure of Blc at 1.8 A resolution using remote SAD with xenon. The structure of Blc, the first of a bacterial lipocalin, exhibits a classical fold formed by a beta-barrel and a alpha-helix similar to that of the moth bilin binding protein. Its empty and open cavity, however, is too narrow to accommodate bilin, while the alkyl chains of two fatty acids or of a phospholipid could be readily modeled inside the cavity. Blc was reported to be expressed under stress conditions such as starvation or high osmolarity, during which the cell envelope suffers and requires maintenance. These data, together with our structural interpretation, suggest a role for Blc in storage or transport of lipids necessary for membrane repair or maintenance. PMID- 15044024 TI - The membrane topology of EmrE - a small multidrug transporter from Escherichia coli. AB - EmrE is a multidrug transporter from Escherichia coli that belongs to the Smr family of small multidrug transporters. The secondary structure of EmrE consists of a four helical bundle, as judged by different techniques. EmrE has been extensively characterized; nevertheless, the membrane topology of EmrE has not been determined yet. Previous work with a homologous Smr protein provided partial information of the membrane topology, however the location of the carboxy terminus remained inconclusive. In this work we probed the membrane topology of EmrE, focusing on the carboxy-terminus of the protein, using two independent approaches. Our results support a secondary structure where the carboxy-terminus faces the cytoplasm, while the first loop faces the periplasm. PMID- 15044025 TI - Lignin dehydrogenative polymerization mechanism: a poplar cell wall peroxidase directly oxidizes polymer lignin and produces in vitro dehydrogenative polymer rich in beta-O-4 linkage. AB - An investigation was performed to determine whether lignin dehydrogenative polymerization proceeds via radical mediation or direct oxidation by peroxidases. It was found that coniferyl alcohol radical transferred quickly to sinapyl alcohol. The transfer to syringaresinol was slower, however, the transfer to polymeric lignols occurred very slightly. This result suggests that the radical mediator theory does not sufficiently explain the mechanism for dehydrogenative polymerization of lignin. A cationic cell wall peroxidase (CWPO-C) from poplar (Populus alba L.) callus showed a strong substrate preference for sinapyl alcohol and the sinapyl alcohol dimer, syringaresinol. Moreover, CWPO-C was capable of oxidizing high-molecular-weight sinapyl alcohol polymers and ferrocytochrome c. Therefore, the CWPO-C characteristics are important to produce polymer lignin. The results suggest that CWPO-C may be a peroxidase isoenzyme responsible for the lignification of plant cell walls. PMID- 15044026 TI - A new approach that allows identification of intron-split peptides from mass spectrometric data in genomic databases. AB - We present a new approach that allows the identification of intron-split peptides from mass spectrometric data in genomic databases. Our algorithm uses small regions of peptide sequence information which are automatically deduced from de novo amino acid sequence predictions together with the molecular mass information of the precursor ion. The sequence predictions are based on selected collision induced mass spectrometric fragmentation spectra. Fragments of the predicted amino acid sequence are aligned with each of the six frames of the translated genome and the precursor mass information is used to assemble the corresponding tryptic peptides using the sequence as a matrix. Hereby, intron-split peptides can be gathered and in turn verified by mass spectrometric data interpretation tools such as Sequest. PMID- 15044027 TI - Eu3+ binding to europium-regenerated bacteriorhodopsin upon delipidation and monomerization. AB - We have studied the effect of monomerization of the purple membrane lattice, as well as removal of 75% of the lipids, on the binding properties of Eu(3+) ions. We found that delipidation and monomerization do not cause the cations to lose their binding ability to the protein. This suggests that the three most strongly bound Eu(3+) cations do not bind to the lipids, but directly bind to the protein. Furthermore, we found that delipidation actually causes a slight increase in the binding affinity. This is likely a result of reduced aggregation of europium regenerated bacteriorhodopsin (bR) upon lipid removal causing more exposure of the binding sites to the Eu(3+) cations. These results, taken with those from our previous publication [Heyes and El-Sayed, Biophys. J. 85 (2003) 426-434], might suggest that the cations remain bound upon delipidation of bR, but have no effect on the function. This is discussed with respect to the role of cations in the function of native bR. PMID- 15044028 TI - Prostaglandins are powerful inducers of NGF and BDNF production in mouse astrocyte cultures. AB - We found that prostaglandin (PG) D(2) and PGE(2), which are major PGs in the brain of mammals, powerfully induced the secretion of nerve growth factor (NGF) from cultured mouse astrocytes; PGE(2) or PGD(2) induced an approximately 12- or 19-fold increase in NGF secretion after a 24-h incubation, respectively. Moreover, it was found that the sequential metabolites of PGD(2), PGJ(2), Delta(12)-PGJ(2), and 15-deoxy-Delta(12,14)-PGJ(2), induced the NGF secretion to the culture medium strikingly (60-98-fold of the control after a 24-h incubation). NGF secretion induced by the J(2) series of PGs was accompanied by the increased expression of NGF mRNA. These PGs also stimulated the secretion/synthesis of brain-derived neurotrophic factor (BDNF). Our findings suggest that PGs play a neuroprotective role by inducing NGF and BDNF production in the central nervous system. PMID- 15044029 TI - Contributions of 5-HT(2C) receptors to multiple actions of central serotonin systems. AB - Insights into neural mechanisms through which central serotonin (5-HT) systems influence brain function may be gained by examining the contributions of individual 5-HT receptor subtypes. Significant attention has focused on the 5 HT(2C) receptor subtype, which is abundantly expressed throughout the central nervous system and displays high-affinity interactions with a wide variety of psychiatric medications. Both pharmacological and genetic approaches to the analysis of 5-HT(2C) receptor function reveal that it contributes substantially to the serotonergic regulation of a wide variety of behavioral and physiological processes. For example, significant inhibitory effects of 5-HT(2C) receptor stimulation have been observed in both limbic and striatal dopamine pathways. These may contribute to the effects of experimental 5-HT(2C) receptor manipulations on responses to psychostimulant, atypical antipsychotic and antidepressant drugs. Further evidence for a role of these receptors in affect regulation arises from recent findings that alterations in 5-HT(2C) mRNA editing are observed in the brains of suicide victims with a history of depression and in animals exposed to antidepressant drug treatment. Finally, we will review a growing body of evidence indicating a role of 5-HT(2C) receptors in the serotonergic regulation of energy balance. Pharmacological and genetic studies reveal these receptors to influence feeding, glucose homeostasis and the energy efficiency of physical activity. PMID- 15044030 TI - Sargaquinoic acid supports the survival of neuronal PC12D cells in a nerve growth factor-independent manner. AB - Sargaquinoic acid (designated previously as MC14) was isolated from a marine brown alga Sargassum macrocarpum, and has been found to possess a novel nerve growth factor (NGF)-dependent neurite outgrowth promoting activity in PC12D cells. In this study, we explored the neuroprotective effects of MC14 in terms of its survival supporting, antioxidant and neurite-regenerating activities under NGF deficient or deprived conditions. Intriguingly, MC14 did not only promote the NGF-induced survival support on neuronal PC12D cells, but also significantly abated neuronal PC12D cell death even in the absence of NGF. The pharmacological inhibition of phosphatidylinositol-3 kinase (PI3K) by wortmannin significantly suppressed the survival supporting activity of MC14, whereas the NGF receptor (tyrosine kinase A or TrkA) inhibitor K252a showed no detectable effect on MC14 activity. These results demonstrate that MC14 supports survival of neuronal PC12D cells in an NGF-independent manner, and that PI3K may be required for the neuroprotective activity of MC14. In addition, we have shown that MC14 markedly enhanced neurite-regeneration and protected PC12D cells from hydrogen peroxide (H(2)O(2))-induced oxidative stress. These pharmacological features suggest that MC14 may be a potentially important neuroprotective agent. PMID- 15044031 TI - Effect of glucocorticoids on 3-nitropropionic acid-induced oxidative stress in synaptosomes. AB - The present study with rat striatal and cortical synaptosomes evaluated the effect of dexamethasone (300 microg/kg i.p./day) with and without simultaneous adrenalectomy on the oxidative stress induced by 3-nitropropionic acid (20 mg/kg/day for 4 days). Adrenalectomy enhanced the oxidative stress induced by 3 nitropropionic acid. These changes were prevented by previous and simultaneous administration of dexamethasone. Adrenalectomy alone induced oxidative stress with decreases in succinate dehydrogenase activity. Our results revealed that adrenal glucocorticoids, and especially dexamethasone (synthetic glucocorticoid), have a protective effect against oxidative stress induced by 3-nitropropionic acid in some brain regions of the Wistar rat. PMID- 15044032 TI - Suppression of cell proliferation with induction of p21 by Cl(-) channel blockers in human leukemic cells. AB - The existence of Cl(-) channels in lymphocytes and neutrophils has been increasingly recognized, but the biological functions are not yet clear. We examined the effects of Cl(-) channel blockers on the cell proliferation and the cell cycle of human leukemic cell lines. The growth of leukemic cells was suppressed most efficiently by NPPB (5-nitro-2-(3-phenylpropylamino) benzoic acid), partially by 9-AC (9-anthracenecarboxylic acid) and tamoxifen, but not by stilbene compounds. NPPB increased the G0/G1 population and induced the expression of p21, one of the critical molecules for G1/S checkpoint. Antisense oligonucleotide for a NPPB-sensitive and stilbene-insensitive Cl(-) channel, ClC 2, sufficiently suppressed the ClC-2 protein synthesis, but did not affect the growth of leukemic cells. These findings suggest that NPPB-sensitive and stilbene insensitive Cl(-) channels other than ClC-2 play important roles in cell cycles and cell proliferation of human leukemic cells. PMID- 15044033 TI - Cyclic GMP-dependent neurite outgrowth by genipin and nerve growth factor in PC12h cells. AB - We have demonstrated previously that a natural iridoid compound, genipin, induced neuritogenesis through activation of nitric oxide synthase (NOS) and mitogen activated protein kinase (MAPK) in PC12h cells. In this paper, we investigated whether cyclic GMP (cGMP) and cGMP-dependent protein kinase (PKG) are involved in the neuritogenesis as a result of NOS activation. Furthermore, we also investigated the relationship between cGMP and MAPK activation in the signaling pathway. The genipin-induced neuritogenesis accompanied by induction of neurofilament was significantly inhibited by 1H-[1,2,4]oxadiazolo[4,3-a] quinoxalin-1-one (ODQ) and KT5823, inhibitors of soluble guanylate cyclase and PKG, respectively. Genipin-induced MAPK phosphorylation was also abolished by ODQ. These inhibitory effects of ODQ were similar to those observed for nerve growth factor (NGF)-induced neurite outgrowth and MAPK phosphorylation. The membrane-permeable cGMP analog, 8-Bromo-cGMP, had prominent neuritogenic activity, which was completely inhibited by a MAPK kinase inhibitor, PD98059. These results suggest that the soluble guanylate cyclase-PKG signaling pathway is important for MAPK activation by genipin as well as NGF during neuritogenesis in PC12h cells. PMID- 15044034 TI - Effect of calcitonin gene-related peptide on angiotensin II-induced proliferation of rat vascular smooth muscle cells. AB - The present study was designed to examine the effect of calcitonin gene-related peptide (CGRP) on angiotensin II-induced proliferation of cultured rat vascular smooth muscle cells. Vascular smooth muscle cells were grown from explants of Sprague-Dawley rat aorta. Vascular smooth muscle cells (between passages 5 and 10) were incubated with 0.1% neonatal calf serum for 48 h, and then treated with angiotensin II (100 nM) in the absence or presence of CGRP for 24 h. The viability, DNA synthesis and cell cycle of vascular smooth muscle cells were measured. Western blotting was used to determine the activity of intracellular extracellular regulated kinase (ERK1/2). Angiotensin II significantly decreased the viability and proliferation of vascular smooth muscle cells, decreased the proliferation index, and increased the activity of ERK1/2; the effects of angiotensin II were inhibited by CGRP (1-100 nM) in a concentration-dependent manner. In conclusion, CGRP significantly inhibits angiotensin II-induced proliferation of vascular smooth muscle cells, an effect related to a decrease in the activation of mitogen-activated protein kinase pathway. PMID- 15044035 TI - Dopamine acutely decreases type 3 Na(+)/H(+) exchanger activity in renal OK cells through the activation of protein kinases A and C signalling cascades. AB - Dopamine D(1)-mediated inhibition of Na(+),K(+)-ATPase activity in opossum kidney (OK) cells involves the sequential activation of the adenylyl cyclase-protein kinase A (PKA) and the phospholipase C-protein kinase C (PKC) pathways. The present study evaluated the signalling cascades involved in dopamine-mediated inhibition of Na(+)/H(+) exchanger isoform 3 (NHE3) in OK cells. The transport kinetics displayed a simple Michaelis-Menten relationship for extracellular Na(+) of 25+/-6 mM. Dopamine and the dopamine D(1)-like receptor agonist SKF 38393 ((+/ )-1-phenyl-2,3,4,5-tetrahydro-(1H)-3-benzazepine-7,8-diol) inhibited NHE3 activity in a concentration-dependent manner; the dopamine D(2)-like receptor agonist quinerolane was devoid of effect. The SKF 38393-mediated inhibition of NHE3 was prevented either by the dopamine D(1)-like receptor antagonist SKF 83566 ((+/-)-7-Bromo-8-8-hydroxy-3 methyl-1-phenyl-2,3,4,5-tetrahydro-1H-3-benzazepine; 1 microM), overnight treatment with cholera toxin (500 ng/ml), the PKA antagonist H-89 (N-(2-[p-bromocinnamylamino]ethyl)-5 isoquinolinesulfonamide hydrochloride; 10 microM), the PKC antagonist chelerythrine (1 microM), or the phospholipase C inhibitor U-73,122 (1-(6-[(17beta]-3-methoxyestra-1,3,5[10]-trien-17-yl) amino] hexyl)-1H-pyrrole-2,5-dione; 3 microM). In addition, dibutyril cAMP (dB-cAMP; 500 microM) was found to increase phospholipase C activity, both in membranes and in cytosol from OK cells; in contrast, phorbol-12,13-dibutyrate (PDB) (1 microM) did not have a significant effect on phospholipase C activity. Pre-treatment of OK cells with the anti-G(s)alpha antibody, but not the anti-G(q/11)alpha antibody, blunted the inhibitory effect of SKF 38393 on NHE3 activity. It is concluded that dopamine D(1)-mediated inhibition of NHE3 in renal OK cells involves both adenylyl cyclase-PKA and the phospholipase C-PKC pathways, a mechanism similar to that described for Na(+),K(+)-ATPase. PMID- 15044036 TI - Mutagenesis at the human tachykinin NK(2) receptor to define the binding site of a novel class of antagonists. AB - The pharmacological profile of novel antagonists endowed with high affinity for the human tachykinin NK(2) receptor is presented. MEN13918 (Ngamma[Nalpha[Nalpha(benzo[b]thiophen-2-yl)carbonyl]-1-aminocyclohexan-1 carboxy]-d-phenylalanyl]-3-cis-aminocyclohexan-1-carboxylic-acid-N-(1S,2R)-2 aminocyclohexyl)amide trifluoroacetate salt) and MEN14268 (Nalpha[Nalpha(benzo[b]thiophen-2-yl)carbonyl)-1-aminocyclopentane-1-carboxyl]-d phenylalanine-N-[3(morpholin-4-yl)propyl]amide trifluoroacetate salt) were more potent in blocking neurokinin A (NKA, His-Lys-Thr-Asp-Ser-Phe-Val-Gly-Leu-Met NH(2)) induced contraction in human, which induced greater contraction in human (pK(B) 9.1 and 8.3) than rat (pK(B) 6.8 and <6) urinary bladder smooth muscle preparation in vitro. In agreement with functional data, in membrane preparations of CHO cells stably expressing the human NK(2) receptors, both MEN13918 and MEN14268 potently inhibited the binding of agonist ([(125)I]NKA, K(i) 0.2 and 2.8 nM) and antagonist ([(3)H]nepadutant, K(i) 0.1 and 2.2 nM, [(3)H]SR48968 K(i) 0.4 and 6.9 nM) radioligands. Using site-directed mutagenesis and radioligands binding we identified six residues in the transmembrane (TM) helices that are critical determinants for the studied antagonists affinity. To visualize these experimental findings, we constructed a homology model based on the X-ray crystal structure of bovine rhodopsin and suggested a possible binding mode of these newly discovered antagonist ligands to the human tackykinin NK(2) receptor. Both MEN13918 and MEN14268 bind amongst TM4 (Cys167Gly), TM5 (Tyr206Ala), TM6 (Tyr266Ala, Phe270Ala), and TM7 (Tyr289Phe, Tyr289Thr). MEN13918 and MEN14268 diverging binding profile at Y289 mutations in TM7 (Tyr289Phe, Tyr289Thr) suggests a relation of their different chemical moieties with this residue. Moreover, the different influence on binding of these two ligands by mutations located deep along the inner side of TM6 (Phe270Ala, Tyr266Ala, Trp263Ala) indicates a nonequivalent positioning, although occupying the same binding crevice. Furthermore, binding data indicate the Ile202Phe mutation, which mimics the wild-type rat NK(2) receptor sequence, as a species selectivity determinant. In summary, data with mutant receptors describe, for these new tachykinin NK(2) receptor antagonists, a binding site which is partially overlapping either with that of the cyclized peptide antagonist nepadutant (cyclo-[[Asn(beta-d-GlcNAc) Asp-Trp-Phe-Dpr-Leu]cyclo(2beta-5beta)] or the nonpeptide antagonist SR48968 ((S) N-methyl-N[4-(4-acetylamino-4-phenylpiperidino)-2-(3,4 dichlorophenyl)butyl]benzamide). PMID- 15044037 TI - Effects of (1R,9S)-beta-hydrastine on l-DOPA-induced cytotoxicity in PC12 cells. AB - (1R,9S)-beta-Hydrastine in lower concentrations of 10-50 microM inhibits dopamine biosynthesis in PC12 cells. In this study, the effects of (1R,9S)-beta-hydrastine on L-DOPA (L-3,4-dihydroxyphenylalanine)-induced cytotoxicity in PC12 cells were investigated. (1R,9S)-Hydrastine at concentrations up to 250 microM did not reduce cell viability. However, at concentrations higher than 500 microM it caused cytotoxicity in PC12 cells, as determined with the 3-(4,5-dimethylthiazol 2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, TUNEL (terminal deoxynucleotidyltransferase dUTP nick-end labeling) method and flow cytometry. Exposure of PC12 cells to cytotoxic concentrations of (1R,9S)-beta-hydrastine (500 and 750 microM) in combination with L-DOPA (20, 50 and 100 microM) after 24 or 48 h resulted in a significant decrease in cell viability compared with the effects of (1R,9S)-beta-hydrastine or L-DOPA alone, and apoptotic cell death was observed. However, the decrease in cell viability induced by (1R,9S)-beta hydrastine was not prevented by the antioxidant N-acetyl-L-cysteine, indicating that it is not mediated by membrane-based oxygen free radical damage. These data suggest that (1R,9S)-beta-hydrastine has a mild cytotoxic effect and at higher concentration ranges aggravates L-DOPA-induced cytotoxicity in PC12 cells. PMID- 15044038 TI - Licofelone, an inhibitor of cyclooxygenase and 5-lipoxygenase, specifically inhibits cyclooxygenase-1-dependent platelet activation. AB - 5-Lipoxygenase/cyclooxygenase inhibitors, possessing anti-inflammatory action and gastric safety due to cyclooxygenase-2 and 5-lipoxygenase inhibition and antiplatelet activity due to cyclooxygenase-1 blockade, would be beneficial in the treatment of ischemic disease because they may reduce, at the same time, inflammation, underlying the atherosclerotic process, and platelet activation, responsible for acute thrombotic events. In this study, we characterized the antiplatelet effects of the new 5-lipoxygenase/cyclooxygenase inhibitor licofelone ([2,2-dimethyl-6-(4-chlorophenyl)-7-phenyl-2,3,dihydro-1H-pyrrolizine 5-yl]-acetic acid. Licofelone completely prevented platelet aggregation induced in platelet-rich plasma by threshold aggregating concentrations of arachidonic acid (0.87+/-0.14 mM) at threshold inhibitory concentrations of 0.75+/-0.35 microM (n=5). Platelet-rich plasma aggregation induced by threshold aggregating concentrations of collagen/adrenalin (0.3+/-0.05 microg/ml and 0.4+/-0.1 microM, respectively) was reduced to 3.2+/-2% of control at licofelone 100 microM, (P<0.05, n=6). Washed platelet aggregation induced by threshold aggregating concentrations of thrombin (0.07+/-0.01 U/ml) was only partially affected by licofelone at concentrations one or two order of magnitude higher than those fully preventing arachidonic acid-induced aggregation (44+/-11% of control at 100 microM, P<0.05, n=7). Failure to prevent aggregation triggered by high concentrations of collagen/adrenalin in aspirin-treated platelets supports cyclooxygenase-1 as a specific target of licofelone. In fact, licofelone inhibited thromboxane B(2) (TxB(2)) production by all the agonists tested at concentrations between 0.5 and 50 microM. At this concentration, TxB(2) production was reduced at values similar to those of unstimulated platelets. These results indicate that, at clinically relevant concentrations, licofelone exerts a potent antiplatelet effect mediated by the inhibition of cyclooxygenase 1 activity. PMID- 15044039 TI - Role of dopamine D1 receptors in the striatal and cortical fos expression induced by the muscarinic agonist pilocarpine. AB - Injections of the muscarinic cholinergic receptor agonist pilocarpine (50 mg/kg) induced pronounced expression of the immediate early gene (IEG) product Fos in the striatum and cortex of rats. Pretreatment with the dopamine D1 receptor antagonist 7-chloro-8-hydroxy-3-methyl-1-phenyl-2,3,4,5-tetrahydro-H-3 benzazepine hydrochloride (SCH-23390; 0.2-2.0 mg/kg) drastically attenuated the pilocarpine response in the striatum, but had no effect in the cortex. In contrast, the muscarinic receptor antagonist scopolamine (0.75-3.00 mg/kg) virtually abolished the Fos response at both sites. These results suggest that stimulation of dopamine D1 receptors may mediate the effects of muscarinic agonists on Fos expression in the striatum, but not the cortex. PMID- 15044040 TI - Spinal antinociceptive effects of AA501, a novel chimeric peptide with opioid receptor agonist and tachykinin receptor antagonist moieties. AB - The use of "multimodal" combination analgesic therapies or novel single molecules possessing multiple analgesic targets is becoming increasingly attractive. In previous experiments we showed that a substance P antagonist injected intrathecally potentiated the antinociceptive effects of potent opioid receptor agonist, biphalin. Based on examination of the biphalin structure-activity relationship, we designed and synthesized a novel chimeric peptide, termed AA501 (N'(Tyr-D-Ala-Gly-Phe), N"(Z-Trp) hydrazide, Z=benzyloxycarbonyl). AA501 consists of an opioid receptor agonist pharmacophore related to biphalin and a substance P receptor antagonist pharmacophore, both linked by a hydrazide bridge. The present study evaluates the ability of a novel chimeric peptide, AA501, to bind to opioid and substance P receptors and to produce antinociception in tail-flick and formalin tests, and in a neuropathic pain model when administered intrathecally to rats. PMID- 15044041 TI - Analogues of homoibotenic acid show potent and selective activity following sensitisation by quisqualic acid. AB - Quisqualic acid induces sensitisation of neurones to depolarisation by analogues of 2-amino-4-phosphonobutyric acid (AP4), phenylglycine, and homoibotenic acid (HIBO). Thus, after administration of quisqualate these analogues become active at concentrations at which they are otherwise inactive. The mechanisms behind quisqualate-induced sensitisation are poorly understood and have not previously been quantified properly. In this study, we have tested the activity of a number of 4-alkyl- and 4-aryl-substituted analogues of HIBO as regards quisqualate sensitisation, and present a method for quantifying the sensitisation induced by quisqualate at cortical neurones. These analogues are generally more potent and selective than (S)-AP4 or its homologue (S)-AP5 following quisqualate sensitisation. Furthermore, we found a statistically significant correlation between the ligands' ability to inhibit CaCl(2)-dependent (S)-[(3)H]glutamate uptake into rat cortical synaptosomes, and their potency following quisqualate induced depolarisation. This demonstrates the involvement of a transport system in the mechanism underlying the quisqualate-effect. PMID- 15044042 TI - Repeated amphetamine treatment causes a persistent elevation of glial fibrillary acidic protein in the caudate-putamen. AB - The ability of repeated D-amphetamine (2 mg/kg) treatment to induce behavioral sensitization in rats and alter glial fibrillary acidic protein (GFAP), dopamine transporter (DAT) and glutamate transporter-1 (GLT-1) immunoreactivities was assessed after a 10-day drug abstinence period. Results showed that a sensitizing regimen of amphetamine caused a persistent increase in the number of GFAP positive cells in the dorsal and ventral caudate-putamen. DAT and GLT-1 immunoreactivities were unaffected. Although the elevated GFAP expression may be due to a mild neurotoxicity, it is also possible that amphetamine-induced increases in GFAP reflect adaptive changes that may be associated with processes underlying behavioral sensitization. PMID- 15044043 TI - alpha,beta-MeATP augments the UTP contraction of rabbit basilar artery. AB - The mechanism underlying the interaction between alpha,beta-methyleneadenosine 5' triphosphate (alpha,beta-MeATP) and uridine 5'-triphosphate (UTP) was investigated using the basilar artery of a rabbit. UTP induced a concentration dependent contraction, whereas P2X receptor agonists, such as alpha,beta-MeATP and 2-methylthioadenosine 5'-triphosphate (2-MeSATP), did not induce any contraction up to 100 microM. alpha,beta-MeATP augmented the UTP contraction two fold, immediately and reversibly. This effect was observed with ectonucleotidase inhibition with 1 mM Ni(2+), the removal of extracellular Ca(2+) or Evans blue. The contractile response to adenosine 5'-O-(3-triphosphate) (ATPgammaS), a selective agonist for P2Y(4), was augmented by pretreatment with alpha,beta-MeATP also. ATPgammaS had no additional effect on the UTP contraction fully activated with alpha,beta-MeATP. UTP (100 microM) did not induce an increase in cytosolic Ca(2+) in a rabbit basilar arterial strip; however, in the presence of 1 mM alpha,beta-MeATP, UTP induced a significant increase in cytosolic Ca(2+). These results suggest that alpha,beta-MeATP facilitates the activation by UTP of the P2Y receptor (P2Y(4)) of the rabbit basilar artery through mechanisms other than nucleotidase inhibition, and that it does not do so via a P2X receptor. PMID- 15044044 TI - In vivo pertussis toxin treatment reduces contraction of rat resistance arteries but not that of mouse trachea. AB - In order to develop an in vitro method for detecting residual pertussis toxin activity in acellular pertussis vaccines, the effects of in vivo pertussis toxin treatment on contraction and relaxation properties of isolated mouse trachea and of isolated rat small mesenteric resistance arteries were studied. In vivo pertussis toxin treatment (24 or 72 microg/kg, intraperitoneally (i.p.)) did not affect contraction and relaxation properties of isolated BALB/c or NIH mouse trachea. In vivo pertussis toxin treatment (30 microg/kg, intravenously) significantly reduced noradrenaline- or KCl-induced maximal contraction and reduced sensitivity to noradrenaline in isolated male Wistar rat small mesenteric resistance arteries. However, in vivo pertussis toxin treatment did not affect relaxation properties of isolated rat small mesenteric resistance arteries. These results support the hypothesis that vasoconstriction-regulating mechanisms and not airway constriction mechanisms are involved in pertussis toxin-induced histamine sensitisation. The vasoconstriction-regulating mechanisms may provide a lead for further development of an in vitro method for measuring biologically active pertussis toxin in acellular pertussis vaccines based on mechanisms involved in the histamine sensitisation test. PMID- 15044045 TI - Mibefradil is more effective than verapamil for restoring post-ischemic function of isolated hearts of guinea pigs with acute renal failure. AB - The deleterious intracellular Ca(2+) overload in the ischemic-reperfusion injury of the heart can be even more expressed in subjects with acute renal failure in whom maintenance of intracellular Ca(2+) has already been disturbed in normoxia. To study the influence of acute renal failure in ischemic-reperfusion injury on the heart, we used isolated Langendorff's hearts of guinea pigs with gentamicin induced acute renal failure. We examined arrhythmias, heart contractility and myocardial cell damage during reperfusion. Two specific Ca(2+) channel antagonists, mibefradil (0.1 and 1 microM) and verapamil (0.1 microM), were used to test the possible involvement of T-type and L-type Ca(2+) channels in these processes. We exposed hearts to 50 min of zero-flow global ischemia and 60 min of reperfusion. During reperfusion, unrecoverable ventricular fibrillation appeared more often in hearts of animals with acute renal failure than in control hearts (80% vs. 0%, respectively). Mibefradil, but not verapamil, applied either pre- or post-ischemically, terminated ventricular fibrillation in all hearts of animals with acute renal failure. Mibefradil (0.1 microM only) improved contractility in hearts of animals with acute renal failure during reperfusion by 30%. During reperfusion, lactate dehydrogenase (LDH) release rate increased less in hearts of guinea pigs with acute renal failure than in control hearts and only verapamil decreased it additionally. Thus, our results suggest a more important role of T- than of L-type Ca(2+) channels in ischemic-reperfusion injury in isolated guinea pig hearts with acute renal failure. PMID- 15044046 TI - Prenatal stress changes rat arterial adrenergic reactivity in a regionally selective manner. AB - A suboptimal fetal environment has been linked to increased risk of cardiovascular disease in adulthood. We investigated whether intrauterine stress (IUS) alters the development of adrenergic reactivity in different types of rat arteries. Intrauterine stress was induced by ligation of the uterine arteries at day 13 of pregnancy in Wistar rats. First-order mesenteric, renal, femoral and saphenous arteries of the 21-day-old male offspring were studied in a myograph. IUS in the rat changes arterial adrenergic reactivity in a regionally selective manner. Adrenoceptor-mediated responses are altered in the renal artery. Maximal contractile responses to phenylephrine were increased, while sensitivity to the alpha(1)-adrenoceptor agonist was decreased. Intrauterine stress significantly reduced contractile responses to norepinephrine and enhanced relaxing responses to isoproterenol in the renal artery. Adrenergic responses were not modified in mesenteric, femoral and saphenous arteries. In the kidneys the densities of [(3)H]prazosin binding sites, periarterial adrenergic nerves and of the glomeruli were not altered after intrauterine stress at day 13 of gestation. The observed regionally selective alterations in arterial reactivity might link a suboptimal fetal environment to the development of cardiovascular disease in the adult. PMID- 15044047 TI - Evidence that alpha(1B)-adrenoceptors are involved in noradrenaline-induced contractions of rat tail artery. AB - The present study characterizes the alpha(1)-adrenoceptor subtypes mediating contractions to noradrenaline in isolated ring preparations of rat tail artery. Concentration-response (E/[A]) curves to noradrenaline were apparently monophasic (pEC(50) 6.47) but became biphasic in the presence of the selective alpha(1A) adrenoceptor antagonist (+/-)-1,3,5-trimethyl-6-[[3-[4-((2,3-dihydro-2 hydroxymethyl)-1,4-benzodioxin-5-yl)-1-piperazinyl]propyl]amino]-2,4(1H,3H) pyrimidinedione (B8805-033). Whereas the first phase of contraction to noradrenaline remained nearly unaffected in the presence of B8805-033 (0.03-3 microM), the second phase was concentration-dependently shifted to the right (pK(B) 8.06). In the presence of B8805-033 (3 microM), noradrenaline-induced contractions (pEC(50) 6.55) were antagonized in a competitive manner by prazosin (pK(B) 9.24), tamsulosin (pK(B) 8.55), 2-(2,6-dimethoxyphenoxyethyl)aminomethyl 1,4-benzodioxane (WB 4101; pK(B) 7.81), spiperone (pK(B) 7.69), 4-amino-2-[4-[1 (benzyloxycarbonyl)-2(S)-[[(1,1-dimethylethyl)amino]carbonyl]-piperazinyl]-6,7 dimethoxyquinazoline (L-765,314; pK(B) 7.31), 5-methylurapidil (pK(B) 6.55), 8-[2 [4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-8-azaspiro[4.5]decane-7,9-dione (BMY 7378; pK(B) 6.43), and 8-[2-(1,4-benzodioxan-2-ylmethylamino)ethyl]-8 azaspiro[4.5]decane-7,9-dione (MDL 73005EF; pK(B) 5.71), and were also antagonized by 100 microM chloroethylclonidine. N-[2-(2 cyclopropylmethoxyphenoxy)ethyl]-5-chloro-alpha,alpha-dimethyl-1H-indole-3 ethanamine (RS-17053) behaved as a noncompetitive antagonist (apparent pA(2) 6.55). Antagonist affinities obtained under these experimental conditions correlated highly with affinities at native and cloned alpha(1B)-adrenoceptors. Pretreatment of arterial rings with B8805-033 (3 microM) followed by receptor inactivation with chloroethylclonidine (100 microM) yielded monophasic E/[A] curves to noradrenaline (pEC(50) 6.14). Noradrenaline-induced contractions were competitively antagonized by tamsulosin (pK(B) 10.32), 5-methylurapidil (pK(B) 8.66), RS-17053 (pK(B) 8.44), B8805-033 (pK(B) 7.87), BMY 7378 (pK(B) 6.54), and L-765,314 (pK(B) 6.41). Antagonist affinities obtained under these experimental conditions correlated highly with affinities at native and cloned alpha(1A) adrenoceptors. It is concluded that the contraction to noradrenaline in rat tail artery is mediated by both alpha(1B)- and alpha(1A)-adrenoceptors, each component of contraction being separable by use of selective alpha(1A)-adrenoceptor blockade and alpha(1B)-adrenoceptor alkylation, respectively. PMID- 15044049 TI - Elastase inhibition reduced death associated with acid aspiration-induced lung injury in hamsters. AB - This study examined whether the specific inhibition of neutrophil elastase by sivelestat sodium hydrate (sivelestat) reduced deaths associated with severe acute lung injury after hydrochloric acid (HCl) aspiration in hamsters. Animals that received a single intratracheal instillation of HCl (0.2 N, 200 microL) time dependently died by occlusion of their trachea with inflammatory exudate. In a time course study, these animals developed severe lung injury, peaking 12 to 24 h after HCl instillation, as indicated by hemorrhage and a massive increase in the protein concentration of bronchoalveolar lavage fluid. These changes were closely correlated with neutrophil elastase activity in bronchoalveolar lavage fluid. Sivelestat (0.01, 0.1 and 1 mg/kg/h), when intravenously infused during the first 48 h post-HCl instillation, dose-dependently reduced death in HCl-instilled hamsters. In a separate experiment, analysis of bronchoalveolar lavage fluid parameters and partial pressure of arterial oxygen (PaO(2)) 8 h post-HCl instillation showed that sivelestat at 1 mg/kg/h, i.v. significantly improved both bronchoalveolar lavage fluid parameters and PaO(2) levels with evidence of the inhibition of neutrophil elastase activity in bronchoalveolar lavage fluid. These results suggest that neutrophil elastase plays a significant role in this type of severe acute lung injury that leads to death by respiratory failure. PMID- 15044048 TI - Nickel inhibits urocortin-induced relaxation in the rat pulmonary artery. AB - Urocortin relaxes rat pulmonary arteries partly through a cyclic AMP-dependent but Ca(2+) channel-independent mechanism. However, other participating mechanisms are relatively unknown. The present study was designed to examine whether the forward mode of Na(+)-Ca(2+) exchangers play a role in the relaxant responses to urocortin in isolated rat small pulmonary arteries. Endothelium-denuded rings were mounted on small vessel myographs for measurement of changes in isometric tension. Urocortin inhibited 9,11-dideoxy-11alpha,9alpha-epoxy methanoprostaglandin F(2alpha) (U46619)-induced contraction in a concentration dependent manner and this inhibition was reversed by astressin, a corticotropin releasing factor receptor antagonist. Micromolar concentrations of nickel (Ni(2+)) chloride, a putative inhibitor of the Na(+)-Ca(2+) exchanger, reduced the relaxant responses to urocortin. Urocortin-induced relaxation was abolished in a Na(+)-free solution, a condition that eliminates influence of the forward mode of Na(+)-Ca(2+) exchanger. In contrast, the relaxant responses to atrial natriuretic peptide or forskolin were unaffected by Ni(2+) or with removal of extracellular Na(+). The present results provide indirect evidence suggesting that stimulation of Na(+)-Ca(2+) exchangers may contribute to urocortin-induced endothelium-independent pulmonary artery relaxation. PMID- 15044050 TI - Bronchoconstriction and endogenous nitric oxide in isolated lungs of spontaneously hypertensive rats. AB - Bronchoconstrictor responses were measured in lungs isolated from spontaneously hypertensive (SHR) and normotensive rats, perfused via the airways. Lungs from SHRs were more responsive than lungs from normotensive rats to methacholine, 5 hydroxytryptamine (5-HT), arachidonic acid or prostaglandin H(2). The responses of SHR airways to methacholine or 5-HT were unaffected by pretreatment in vivo with an inhibitor of nitric oxide (NO) synthase, N(omega)-nitro-L-arginine methyl ester hydrochloride (L-NAME, 30 mg kg(-1)), although responses in normotensive airways to methacholine, but not to 5-HT, were enhanced. Antigen challenge of isolated lungs from actively sensitized rats elicited bronchoconstriction, not different between strains. Pretreatment with L-NAME increased the response to antigen challenge only in normotensive lungs. Compound 48/80 induced bronchoconstriction in lungs from either strain, equally. These responses to compound 48/80 were unaffected by L-NAME pretreatment. Thus, SHR airways lack relaxing factors and degranulation of mast cells in SHR lungs was not affected by endogenous NO. PMID- 15044051 TI - Mechanisms responsible for the in vitro relaxation of a novel dibenzothiepine derivative (NSU-242) on tracheal and vascular smooth muscles. AB - In our previous general screening experiments, we found that NSU-242, a dibenzothiepine derivative (1-10 mg/kg), inhibited antigen-induced immediate asthmatic response in actively sensitized guinea pigs in a dose-dependent manner. The purpose of the present study was to assess the mechanism of the relaxing effect of NSU-242 on smooth muscle contractions in isolated smooth muscle tissues of the porcine trachea and rat aorta. NSU-242 administration resulted in a concentration-dependent inhibition of the tracheal-tissue contractions induced by carbachol and high K(+) and the aortic-tissue contractions induced by norepinephrine and high K(+). The IC(50) values of these inhibitions were 1-10 microM, and there was no selectivity for the type of stimulation. In tracheal tissue, the relaxations were accompanied by neither changes in cAMP nor changes in cGMP. Carbachol (1 microM) and high K(+) (59.2 mM) increased myosin light chain (MLC) phosphorylation in the trachea, and NSU-242 (3-30 microM) had no effect on the level of MLC phosphorylation. Furthermore, NSU-242 (300 microM) had no effect on contractions in membrane-permeabilized tracheal tissue. FITC phalloidin staining of the actin fiber in cultured vascular smooth muscle cells (A7r5) indicated that NSU-242 (10-100 microM) altered the configuration of actin stress fiber in the cytosol. However, unlike cytochalasin D, NSU-242 did not inhibit actin polymerization as assessed by in vitro assay. These results suggest that NSU-242 inhibits smooth muscle contractions without any effect on the Ca(2+) dependent MLC phosphorylation. NSU-242 may uncouple the force generated by the activated actomyosin interaction, possibly by modifying the actin assembly in smooth muscle cells without a direct effect on actin molecules. PMID- 15044052 TI - Central and peripheral mechanisms contribute to the antiemetic actions of delta-9 tetrahydrocannabinol against 5-hydroxytryptophan-induced emesis. AB - Delta-9-tetrahydrocannabinol (delta-9-THC) prevents cisplatin-induced emesis via cannabinoid CB(1) receptors. Whether central and/or peripheral cannabinoid CB(1) receptors account for the antiemetic action(s) of delta-9-THC remains to be investigated. The 5-hydroxytryptamine (5-HT=serotonin) precursor, 5 hydroxytryptophan (5-HTP), is an indirect 5-HT agonist and simultaneously produces the head-twitch response (a centrally mediated serotonin 5-HT(2A) receptor-induced behavior) and emesis (a serotonin 5-HT(3) receptor-induced response, mediated by both peripheral and central mechanisms) in the least shrew (Cryptotis parva). The peripheral amino acid decarboxylase inhibitor, carbidopa, prevents the conversion of 5-HTP to 5-HT in the periphery and elevates 5-HTP levels in the central nervous system (CNS). When administered i.p. alone, a 50 mg/kg dose of 5-HTP failed to induce either behaviour while its 100 mg/kg dose produced robust frequencies of both head-twitch response and emesis. Pretreatment with carbidopa (0, 10, 20 and 40 mg/kg) potentiated the ability of both doses of 5-HTP to produce the head-twitch response in a dose-dependent but bell-shaped manner, with maximal potentiation occurring at 20 mg/kg carbidopa. Carbidopa dose dependently reduced the frequency of 5-HTP (100 mg/kg)-induced emesis, whereas the 10 mg/kg dose potentiated, and the 20 and 40 mg/kg doses suppressed the frequency of vomits produced by the 50 mg/kg dose of 5-HTP. The peripheral and/or central antiemetic action(s) of delta-9-THC (0, 1, 2.5, 5, 10 and 20 mg/kg) against 5-HTP (100 mg/kg)-induced head-twitch response and emesis were investigated in different groups of carbidopa (0, 10 and 20 mg/kg) pretreated shrews. Irrespective of carbidopa treatment, delta-9-THC attenuated the frequency of 5-HTP-induced head-twitch response in a dose-dependent manner with similar ID(50) values. Although delta-9-THC also reduced the frequency of 5-HTP-induced emesis with similar ID(50s), at the 5 mg/kg delta-9-THC dose however, 5-HTP induced significantly less vomits in the 10 and 20 mg/kg carbidopa-treated groups relative to its 0 mg/kg control group. Moreover, increasing doses of carbidopa significantly shifted the inhibitory dose-response effect of delta-9-THC in protecting shrews from 5-HTP-induced emesis to the left. Relatively, a large dose of delta-9-THC (20 mg/kg) was required to significantly reduce the number of vomits produced by direct acting serotonergic 5-HT(3) receptor agonists, serotonin and 2-methylserotonin. Low doses of delta-9-THC (0.1-1 mg/kg) nearly completely prevented 2-methylserotonin-induced, centrally mediated, head-twitch and ear-scratch responses. The results indicate that delta-9-THC probably acts pre- and postsynaptically to attenuate emesis produced by indirect and direct acting 5-HT(3) receptor agonists via both central and peripheral mechanisms. In addition, delta-9-THC prevents 5-HTP-induced head-twitch and emesis via cannabinoid CB(1) receptors since the CB(1) receptor antagonist, SR 141716A [N piperidino-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methylpyrazole-3 carboxamide], countered the inhibitory actions of an effective dose of delta-9 THC against both behaviours. PMID- 15044053 TI - The combination of metformin and a dipeptidyl peptidase IV inhibitor prevents 5 fluorouracil-induced reduction of small intestine weight. AB - Glucagon-like peptide 2 (GLP-2), which has intestinotrophic effects, is secreted from L-cells in the intestine in response to nutrient ingestion and is degraded by dipeptidyl peptidase IV (DPPIV). In this report, we show that biguanides promote GLP-2 release. Plasma GLP-2 levels were significantly increased by 1.4- to 1.6-fold in fasted F344 rats 1 h after oral meformin (300 mg/kg), phenformin (30 and 100 mg/kg) and buformin (100 mg/kg) treatment. In addition, metformin administration (300 mg/kg, p.o.) significantly elevated plasma GLP-2 in fasted CD 1 mice by about 2.0-fold 1 and 3 h after the treatment. Metformin and/or valine pyrrolidide, a DPPIV inhibitor, was orally given (300 and 30 mg/kg, respectively, p.o., b.i.d., 3 days) to BALB/c mice treated with 5-fluorouracil (5-FU; 60 mg/kg, s.i.d.), which induces gastrointestinal damage leading to a reduction of small intestine wet weight. Metformin and valine-pyrrolidide co-administration prevented the 5-FU-induced reduction of wet weight of the small intestine, whereas metformin or valine-pyrrolidide alone had no effect. These results suggest that GLP-2 is co-secreted with GLP-1 flollowing biguanide stimulation, and that the combination of metformin with a DPPIV inhibitor might a useful oral treatment for gastrointestinal damage, based on GLP-2 actions. PMID- 15044054 TI - Effect of adrenomedullin on c-Met receptor expression after reserpine-induced gastric damage in the rat. AB - Here, we show an increase in c-Met receptor expression during reserpine-induced gastric damage in the rat, as assessed by immunohistochemistry. Pretreatment of animals with adrenomedullin prevented this increase in c-Met expression. c-Met immunoreactivity was localized in gastric glands. c-Met immunoreactivity was associated with increased phosphorylation of c-Met receptor and extracellular signal-regulated kinase (ERK(1/2)). Our results suggest that both adrenomedullin and c-Met act as parallel defence mechanisms during pharmacologically induced gastric mucosa injury. PMID- 15044055 TI - A new pyrazolo pyrimidine derivative inhibitor of cyclooxygenase-2 with anti angiogenic activity. AB - In a previous study, we reported a new pyrazolo pyrimidine derivative, N(4) benzyl-N(6),N(6)-dimethyl-1-1(tert-butyl)-1H-pyrazolo[3,4-d]pyrimidine-6,4 diamine (DPP), which inhibited potently cyclooxygenase-2 activity in intact cell assays with minor activity against cyclooxygenase-1 (IC(50)=0.9 nM for cyclooxygenase-2 versus IC(50)=59.6 nM for cyclooxygenase-1). In the present work, this behaviour was confirmed in vivo by using the 24-h zymosan-injected mouse air pouch model (ID(50)=1.36 nM/pouch for prostaglandin E(2) level). We also studied the possible beneficial effect of DPP in the angiogenesis-dependent murine air pouch granuloma and rat paw carrageenan-induced hyperalgesia models. DPP exerted analgesic and anti-angiogenic (52% reduction in angiogenesis at 10 mg/kg, i.p.) effects that may be associated with inhibition of cyclooxygenase-2 activity. PMID- 15044056 TI - JTE-607, a cytokine release blocker, attenuates acid aspiration-induced lung injury in rats. AB - The present study was designed to clarify the effects of (-)-ethyl N-[3,5 dichloro-2-hydroxy-4-[2-(4-methyl-piperazin-1-yl)ethoxy]benzoyl]-l phenylalaninate dihydrochloride (JTE-607), a novel multiple cytokine inhibitor, on hydrochloric acid (HCl) aspiration lung injury in rats. HCl (0.1 N, 2 ml kg( 1)) was instilled into male Sprague-Dawley rats that were pretreated with or without JTE-607 (30 or 75 mg kg(-1) h(-1)). As a control, normal saline (2 ml kg( 1)) was instilled in rats. All the animals were anesthetized with intraperitoneally injected pentobarbital sodium (40 mg kg(-1)). Bronchoalveolar lavage was performed 5 h (h) after HCl or normal saline instillation. In bronchoalveolar lavage fluid, the increases in total nuclear cell counts, neutrophil counts, optical density at 412 nm as an indication of pulmonary hemorrhage, concentrations of albumin, tumor necrosis factor (TNF)-alpha, interleukin (IL)-6 and cytokine-induced neutrophil chemoattractant induced by HCl instillation were significantly reduced by JTE-607 pretreatment. The level of expression of tumor necrosis factor-alpha and interleukin-6 mRNA in lung tissue was analyzed. The mean expression level of tumor necrosis factor-alpha and interleukin-6 mRNA in the JTE-607 group was lower than that in the HCl and NS groups. The wet-to-dry weight ratio was also determined, and JTE-607 at the dose of 75 mg kg(-1) h(-1) significantly attenuated the increased wet-to-dry weight ratio induced by HCl. These results suggest that JTE-607 can inhibit the production of inflammatory cytokines such as tumor necrosis factor-alpha, interleukin-6 and cytokine-induced neutrophil chemoattractant and attenuate acid induced lung injury in rats. This agent might be therapeutically useful for lung injury. PMID- 15044057 TI - Chronopharmacology of oxacalcitriol in rat model of osteoporosis. AB - We have previously reported the merits of chronopharmacological effect of 1 alpha(OH) vitamin D3 in aged stroke-prone spontaneously hypertensive rat (SHRSP), a model of osteoporosis [Eur. J. Pharmacol. 428 (2001) 283.]. In this study, the chronopharmacological effect of 22-oxacalcitriol, a newly developed active vitamin D3 analogue with less calcemic activity, was evaluated by a single and repeated dosing of the drug in aged SHRSP. Animals (7 months old) were kept in rooms with a 12-h light/dark cycle. Single (12.5 microg/kg, i.v.) and repeated (5 microg/kg, i.v. three times a week for 12 weeks) dosing of 22-oxacalcitriol or vehicle was given at either 2 h after lights on (2HALO) or 14 h after lights on (14HALO). The severity of adverse reactions such as the changes of body weight, hypercalcemia and hyperphosphatemia, was significantly mild when the drug was given at 14HALO. Especially, the increase of serum Ca concentration was not detected at 14HALO trial. Serum concentrations of total (protein-bound and unbound) 22-oxacalcitriol and albumin (a major binding protein of the drug) of the 2HALO and 14HALO trials did not significantly differ. The decrease of parathyroid hormone (PTH) concentration was greater in the 14HALO trial while the increase in urinary ratio of Ca to creatinine was greater in the 2HALO trial. The increase in bone density of both femurs at the end of the study was greater in the 14HALO trial. The suppression of urinary excretion of deoxypyridinoline, an index of bone resorption capacity of osteoclast, was greater in the 14HALO trial, which indicates that the efficacy of 22-oxacalcitriol for suppressing bone resorption might vary with the dosing time. This is the first study to show the dosing-time-dependent changes in the efficacy and toxicity of 22-oxacalcitriol in the animal model of osteoporosis. Chronopharmacological differences seem to be more prominent than those of other vitamin D analogues. To use 22-oxacalcitriol at an adequate timing might provide better efficacy and safety than other vitamin D3 analogues for the treatment of osteoporosis. PMID- 15044058 TI - Postsynaptic alpha 4 beta 2 and alpha 7 type nicotinic acetylcholine receptors contribute to the local and endogenous acetylcholine-mediated synaptic transmissions in nigral dopaminergic neurons. AB - The local and endogenous nicotinic neuronal transmissions of dopaminergic neurons in the substantia nigra were confirmed electrophysiologically using a slice-patch technique. After identifying dopaminergic neurons based on their electrophysiological characteristics, miniature postsynaptic inward currents were recorded in the presence of atropine (a muscarinic acetylcholine receptor antagonist), bicuculline (a GABA receptor antagonist) and L-glutamic acid diethyl ester (GDEE) (a non-selective glutamate receptor antagonist). Under conditions that eliminated muscarinic, GABAergic and glutamatergic synaptic transmissions, we found miniature currents that were inhibited by the specific neuronal nicotinic receptor antagonists, dihydro-beta-erythroidine (DHbetaE) and/or methyllycaconitine (MLA) (selective alpha4beta2 and/or alpha7 nicotinic acetylcholine receptor antagonists, respectively). Under the same extracellular conditions, local stimulations in the vicinity of a target neuron evoked excitatory postsynaptic inward currents (EPSCs). These EPSCs were elicited in an extracellular Ca(2+) dependent manner and were also blocked by DHbetaE and/or MLA. These results suggest that dopaminergic neurons in the substantia nigra receive excitatory cholinergic inputs that are mediated via at least two types of postsynaptic nicotinic receptors, namely alpha7 and alpha4beta2 subtypes. PMID- 15044059 TI - Presence and role of nitric oxide in the central nervous system of the freshwater snail Planorbarius corneus: possible implication in neuron-microglia communication. AB - The aim of the present study was to investigate the involvement of nitric oxide (NO) as a messenger molecule in neuron-microglia communication in the central nervous system (CNS) of the freshwater snail Planorbarius corneus. The presence of both neuronal (nNOS) and inducible nitric oxide synthase (iNOS) was studied using NADPH-diaphorase (NADPH-d) histochemistry and NOS immunocytochemistry. The experiments were performed on whole ganglia and cultured microglial cells after different activation modalities, such as treatment with lipopolysaccharide and adenosine triphosphate and/or maintaining ganglia in culture medium till 7 days. In sections, nNOS immunoreactivity was found only in neurons and nNOS-positive elements were less numerous than NADPH-d-positive ones, with which they partially overlapped. The iNOS immunoreactivity was observed only after activation, in both nerve and microglial cells. We also found that the number of iNOS-immunoreactive neurons and microglia varied, depending on the activation modalities. In microglial cell cultures, iNOS was expressed in the first generation of cells only after activation, whereas a second generation, proliferated after ganglia activation, expressed iNOS even in the unstimulated condition. PMID- 15044060 TI - Combination therapy of stroke in rats with a nitric oxide donor and human bone marrow stromal cells enhances angiogenesis and neurogenesis. AB - We tested the hypothesis that intravenous infusion of human marrow stromal cells (hMSC) with a nitric oxide donor, (Z)-1-[N-(2-aminoethyl)-N-(2-ammonioethyl) aminio] diazen-1-ium-1,2-diolate (DETA/NONOate), enhances angiogenesis, neurogenesis and neurological functional recovery after stroke in rats compared to individual therapy. Experimental groups consist of rats subjected to 2 h of middle cerebral artery occlusion (MCAo) and at 24 h after MCAo intravenous injection of (n=10/group): Group 1: phosphate buffered saline (PBS 1 ml) for control. Group 2: NONOate alone (0.4 mg/kg). Group 3: hMSCs (1 x 10(6)) alone. Group 4: hMSCs (1 x 10(6)) with NONOate (0.4 mg/kg). Functional tests and immunohistochemical staining were performed. Marginal functional recovery after treatment of stroke was found with 1 x 10(6) hMSCs alone (p=0.06) and no benefit was detected with NONOate alone (0.4 mg/kg, p=0.64). However, NONOate+hMSCs in combination significantly induced functional recovery (p<0.05). Treatment using hMSC in combination with NONOate significantly increased vessel perimeter and endothelial cell proliferation compared with hMSC or NONOate alone treatment (p<0.05). Cell proliferation and neurogenesis were assessed with bromodeoxyuridine (BrdU) labeling and immunostaining for cell type-specific markers. Combination treatment promoted increased, BrdU positive cell number in the subventricular zone (SVZ), migrating neuronal doublecortin immunoreactive cells and VEGF and bFGF expression in the ischemic boundary area compared to individual treatment. The functional therapeutic enhancement of combination treatment may be attributed to increased plasticity induced by the combination of a nitric oxide donor and hMSC therapy. These data suggest that pharmacological and cellular therapy may provide an additive therapeutic benefit after stroke. PMID- 15044061 TI - Early exploratory behavior abnormalities in R6/1 Huntington's disease transgenic mice. AB - The Huntington's disease (HD) R6/1 transgenic mouse model, containing a human huntington gene exon-1 with approximately 115 CAG repeats, has multiple biochemical and neuroanatomical abnormalities. Overt neurological symptoms have a relatively late onset (15-21 weeks of age). In this paper, we report exploratory behavior abnormalities that appear well before the onset of obvious pathology. The first differences in exploratory behaviors were evident by 4 weeks of age, when R6/1 mice were hyperactive relative to wild-type controls. However, by 6-7 weeks of age, R6/1 mice were less active than controls. R6/1 mice traveled less in the activity monitor, engaged in fewer stereotypic movements, spent more time resting, and traveled less distance per movement than did wild-type controls. R6/1 mice also displayed intersession habituation abnormalities over the 3 days of testing. These behavioral abnormalities precede the earliest neurochemical and molecular changes reported in the literature to date, and thus indicate subtle early pathology that has not yet been documented. These behavioral abnormalities also occur prior to weight loss in the transgenic mice. Since we were able to detect an abnormal phenotype at an early age in R6/1 mice, this assay may be a useful tool for evaluating therapeutic agents. PMID- 15044062 TI - Ca2+/calmodulin-dependent protein kinase II in the rat cranial sensory ganglia. AB - Immunohistochemistry for Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) was performed on the rat cranial sensory ganglia. More than one half of neurons was immunoreactive for the enzyme in the trigeminal (60%), jugular (70%), petrosal (55%) and nodose ganglia (63%). These neurons were mainly small to medium-sized. The co-expression study demonstrated that one half of CaMKII immunoreactive (ir) neurons was also immunoreactive for calcitonin gene-related peptide (CGRP) or the vanilloid receptor subtype 1 (VR1) in the trigeminal, jugular and petrosal ganglia. In the nodose ganglion, CaMKII-ir neurons were mostly devoid of CGRP-immunoreactivity (ir) (8.2%) whereas the co-expression with VR1-ir was common among such neurons (72%). In the facial skin, nasal mucosa and palate, the epithelium and taste bud were innervated by CaMKII-ir nerve fibers. In addition, the retrograde tracing study demonstrated that 39.6% and 44.8% of trigeminal neurons which were retrogradely traced with fluorogold from the facial skin and nasal mucosa exhibited CaMKII-ir. Forty-six percent of petrosal neurons which innervated the soft palate were immunoreactive for the enzyme. PMID- 15044063 TI - PSD-95 promotes CaMKII-catalyzed serine phosphorylation of the synaptic RAS GTPase activating protein SynGAP after transient brain ischemia in rat hippocampus. AB - Recent studies have indicated that cerebral ischemia induces rapid serine phosphorylation of synaptic RAS-GTPase activating protein (SynGAP) by calcium/Camodulin-dependent protein kinase II (CaMKII) in rat hippocampus. To further illustrate the mechanisms underlying these processes, we examined the effects of transient (15 min) brain ischemia followed by reperfusion (0, 30 min, 6 h, 1, 3 days) on serine phosphorylation of SynGAP and interactions involving SynGAP, postsynaptic density protein 95 (PSD95) and CaMKII in rat hippocampus. Transient brain ischemia was induced by the method of four-vessel occlusion in Sprague-Dawley rats. Serine phosphorylation of SynGAP increased immediately after brain ischemia and peaked at 30-min reperfusion, and the increase was maintained for 3 days. The association among SynGAP, PSD95 and CaMKII had a similar trend as serine phosphorylation of SynGAP. Intracrebroventricular infusion of PSD95 antisense oligodeoxynucleotide not only markedly decreased the protein levels of PSD95 but also attenuated the elevated serine phosphorylation of SynGAP and the associations among SynGAP, PSD95 and CaMKII induced by 30-min reperfusion following 15-min brain ischemia. The results suggest that the serine phosphorylation of SynGAP catalyzed by CaMKII is immediately increased and that PSD95 is critical for promoting SynGAP serine phosphorylation after transient brain ischemia. PMID- 15044064 TI - Properties of native P2X receptors in large multipolar neurons dissociated from rat hypothalamic arcuate nucleus. AB - ATP, the ligand of P2X receptors, is a candidate of neurotransmitter or co transmitter in the peripheral and the central nervous systems. Anatomical studies have revealed the wide distribution of P2X receptors in the brain. So far, P2X mediated small synaptic responses have been recorded in some brain regions. To determine the physiological significance of postsynaptic ATP receptors in the brain, we have investigated the P2X responses in rat dissociated hypothalamic arcuate neurons by using the patch-clamp technique. ATP evoked inward currents in a concentration-dependent manner (EC(50)=42 microM) at a holding potential of -70 mV. The current-voltage relationship showed a marked inward rectification starting around -10 mV. Although neither 300 microM alphabeta-methylene-ATP nor 300 microM betagamma-methylene-ATP induced any currents, 100 microM ATPgammaS and 100 microM 2-methylthio-ATP evoked inward currents of which amplitude was about 60% of the control currents evoked by 100 microM ATP. PPADS, one of P2 receptor antagonists, inhibited the ATP-evoked currents in a time- and a concentration dependent manners (IC(50)=19 microM at 2 min). Permeant Ca(2+) inhibited the ATP evoked currents in the range of millimolars (IC(50)=7 mM); however, Cd(2+) (1-300 microM), a broad cation channel blocker, facilitated the currents with slow off response. Zn(2+) in the range of 1-100 microM facilitated the currents whereas Zn(2+) at the concentrations over 100 microM inhibited the currents. These observations suggest that functional P2X receptors are expressed in the hypothalamic arcuate nucleus. The most likely subunit combinations of the P2X receptors are P2X(2)-homomultimer and P2X(2)/P2X(6)-heteromultimer. PMID- 15044065 TI - Ion channels in epithelial cells of the choroid plexus isolated from the lateral ventricle of rat brain. AB - Whole-cell patch clamp methods were used to determine the expression of ion channels in the epithelial cells of choroid plexus isolated from the lateral ventricle of the rat brain. A single population of cells with a mean capacitance of 61.5+/-1.7 pF was identified in 103 recordings. This value is significantly greater than that measured for cells from the fourth ventricle (P<0.01 by unpaired t-test), indicating that cells from the lateral ventricle have a greater surface area. Voltage-dependent, outward currents were recorded using a K(+)-rich electrode solution. These currents were partially inhibited by 10 nM margatoxin or 10 nM dendrotoxin-K and blocked by 5 mM TEA(+). An inward-rectifying chloride conductance was observed in K(+)-free solutions. The relative permeability of this conductance to anions was P(I)>P(Cl)>P(aspartate). A volume-sensitive anion conductance was observed when cell swelling was induced using a hypertonic electrode solution. The properties of each conductance were similar to conductances previously identified in fourth ventricle choroid plexus cells. Furthermore, there were no significant differences between the magnitudes of any of the conductances in cells from the lateral and fourth ventricle choroid plexus. Thus, the ionic conductances expressed in rat lateral and fourth ventricle choroid plexus are very similar. PMID- 15044066 TI - Salsolinol, a dopamine-derived tetrahydroisoquinoline, induces cell death by causing oxidative stress in dopaminergic SH-SY5Y cells, and the said effect is attenuated by metallothionein. AB - The endogenous neurotoxin, 1-methyl-6,7-dihydroxy-1,2,3,4-tetrahydroisoquinoline (salsolinol), has been considered a potential neurotoxin in the etiology of Parkinson's disease (PD). Salsolinol and N-methyl(R)-salsolinol were identified in the brains and cerebrospinal fluid (CSF) of PD patients. Oxidative stress is known to be one of the major contributing factors in the cascade that may finally leads to the cell death in PD. The present study was undertaken to understand the role of salsolinol in oxidative-mediated neuronal toxicity in dopaminergic SH SY5Y cells, and the neuroprotective effects of metallothionein (MT) against salsolinol toxicity in MT overexpressing (MT(trans)) fetal mesencephalic cells. Salsolinol increased the production of reactive oxygen species (ROS) and significantly decreased glutathione (GSH) levels and cell viability in SH-SY5Y cells. Salsolinol also decreased intracellular ATP levels and induced nuclear condensation in these cells. Salsolinol-induced depletion in cell viability was completely prevented by N-acetylcysteine in SH-SY5Y cells, and also prevented by MT in MT(trans) fetal mesencephalic cells compared to control(wt) cells. The extent of nuclear condensation and caspase activation was also less in MT(trans) cells than control(wt) cells. These results suggest that salsolinol causes oxidative stress by decreasing the levels of GSH and by increasing ROS production, and these events may lead to the death of dopaminergic cell. Furthermore, MT overexpression may protect dopaminergic neurons against salsolinol-induced neurotoxicity, most probably by the inhibition of oxidative stress and apoptotic pathways including caspase-3 activation. PMID- 15044067 TI - Vasodilation of brain surface arterioles by blockade of Na-H+ antiport and its inhibition by inhibitors of KATP channel openers. AB - Pial artrioles of rats were monitored in vivo and found to dilate in dose dependent fashion upon application of either benzamil or ethyl isopropyl amiloride, both of which are inhibitors of the sodium-hydrogen antiport. Antiport blockade is known to decrease the internal pH of vascular smooth muscle (VSM). The dilation was blocked by 1 microm glibenclamide, which in that dose is a selective inhibitor of ATP sensitive potassium channels (K(ATP)). The nitric oxide synthase inhibitor nitro-l arginine (l-NNA) also blocked the response. Previous studies of this preparation under the same experimental conditions showed that l-NNA inhibited dilation by K(ATP) openers and that nitric oxide had no permissive action in this setting. Moreover, one study by others has demonstrated a pH sensitive site on the internal surface of K(ATP) while another study by others has demonstrated that sodium propionate, a direct acidifier of the cell, dilates rat basilar artery in K(ATP)-dependent fashion. Therefore, the present data support the following conclusions: decrease of internal pH dilates brain arterioles; the response is K(ATP) dependent; in some situations, inhibitors of nitric oxide synthase can inhibit K(ATP) and K(ATP)-dependent dilations including those produced by decrease of internal pH. PMID- 15044068 TI - Inhibition of glycogen synthase kinase 3 beta is involved in the resistance to oxidative stress in neuronal HT22 cells. AB - Oxidative stress is involved in several neurodegenerative diseases including Alzheimer's disease, Parkinson's disease and ischemic reperfusion injury (stroke). We have established clones of the murine hippocampal neuronal cell line HT22, which are resistant to the oxidative stress-causing agents glutamate and hydrogen peroxide, respectively. These cell clones show a mutual cross-resistance to other oxidative stressors, but not to essentially non-oxidative neurotoxins. We have discovered that the amount of phosphorylated, inactive glycogen synthase kinase (GSK) 3beta is elevated in both resistant clones. Pharmacological inhibition of GSK-3beta with lithium chloride in the sensitive parental neuronal cells results in an increased tolerance to glutamate and hydrogen peroxide, suggesting that GSK-3beta is involved in the control of oxidative stress resistance in these cells. PMID- 15044069 TI - The role of glutathione in nitric oxide donor toxicity to SN56 cholinergic neuron like cells. AB - Our study was designed to determine if compounds used experimentally to generate nitric oxide excess differ in ability to elicit degenerative stress to cholinergic neurons and, if so, what mechanisms account for their differences. Nitric oxide donors are often used experimentally in attempts to emulate the bioactivities of endogenous NO, but the pharmacological actions of NO donors can vary dramatically according to the species of NO (NOx) and other agents (e.g., iron cations, cyanide anion, superoxide anion) released, and as affected by the state of the cellular redox environment. To determine whether different types of NO donors exert differential toxicity in a cholinergic neuronal model, we measured cell viability markers, indicators of NOx formation, levels of intracellular-reduced glutathione (GSH), protein nitrosothiols, and the activation of the transcription factor NF-kappaB in a mouse medial septal cholinergic cell line (clone SN56) following exposure to the NO donors S-nitroso N-acetyl-dl-penicillamine (SNAP), 3-morpholinosydnonimine (SIN-1), or sodium nitroprusside (SNP). SNAP and SIN-1, but not SNP, elicited dramatic increases in media nitrite and intracellular NOx-related fluorescence from cells preloaded with a NOx indicator. Nevertheless, SN56 cells were readily killed by SNP (IC(50) approximately 0.5 mM), while even higher levels (up to 2 mM) of SNAP or SIN-1 were essentially ineffective. SNAP (an NO(+) generator) and SIN-1 (a peroxynitrite generator) both caused increases in SN56 GSH levels; in contrast, SNP caused an immediate and rapid decline in GSH. The increase in GSH in response to SNAP and SIN-1 probably indicates augmentation of intracellular defense mechanisms, because prior depletion of GSH rendered the cells vulnerable to these two donors. GSH depletion did not change the potency of SNP, but GSH depletion made SNAP about twice as potent as SNP. SNAP and SNP, but not SIN-1, activated the transcription factor NF-kappaB, as indicated by increases in p65 nuclear immunoreactivity. Treatment with SNAP, but not SNP or SIN-1, increased levels of S-nitrosothiols in SN56 proteins, consistent with the transfer of an NO(+) equivalent to intracellular thiols. Our experiments show that these three NO donors differ dramatically in their ability to intoxicate SN56 cells, probably because of the different species of NOx and other agents they release, and as reflected in their differing modes of interaction with cellular antioxidant and survival systems. PMID- 15044070 TI - Brain temperature fluctuations during passive vs. active cocaine administration: clues for understanding the pharmacological determination of drug-taking behavior. AB - While it is generally assumed that cocaine self-administration (SA) is determined and maintained by the pharmacological actions of cocaine in the brain, it is also a drug-motivated and drug-reinforced goal-directed behavior, which is determined by concurrent learning and behavioral performance. To dissociate the contributions of pharmacological and behavioral factors to cocaine SA, it is important to compare cocaine SA with its pharmacological copy, passive intravenous (iv) cocaine administration. This approach was employed in the present study with respect to brain temperatures, a dynamic parameter that reflects metabolic neural activity and shows consistent fluctuations during cocaine SA. Passive cocaine injections performed with the same dose/pattern as SA induced brain temperature fluctuations similar in many ways to those in behaving animals. The initial passive drug administration of a session elevated brain temperature, while subsequent repeated injections were associated with biphasic temperature fluctuations that maintained at a relatively stable plateau. Although the magnitude of these fluctuations was twofold smaller than in behaving animals, passive animals had the same pattern; brain temperatures transiently decreased after cocaine injection, then increased, and were inhibited again by the next cocaine infusion. In contrast to self-administering animals, rats exposed to passive cocaine injections had significantly lower basal temperatures and never showed gradual temperature increases preceding the initial injection. Striking differences in brain temperature dynamics seen in the beginning of a session suggest that during the development of drug-taking behavior the initial cocaine induced neural activation becomes transformed into behavior-related "anticipatory" neural activation (motivational arousal) that fuels drug seeking and results in the initial drug intake. While this activation is triggered by drug-related cues and enhanced by the initial cocaine intake, subsequent highly cyclical cocaine intakes appear to be primarily pharmacologically determined. PMID- 15044071 TI - Cannabinoids reduce cAMP levels in the striatum of freely moving rats: an in vivo microdialysis study. AB - The cannabinoid receptor subtype 1 (CB1R) is a member of the G(i)-protein-coupled receptor family and cannabinoid signaling is largely dependent on the suppression of adenylyl cyclase-catalyzed cAMP production. In cell lines transfected with the CB1R or in native tissue preparations, treatment with cannabinoid agonists reduces both basal and forskolin-stimulated cAMP synthesis. We measured extracellular cAMP concentrations in the striatum of freely moving rats utilizing microdialysis to determine if changes in cAMP concentrations in response to CB1R agonists can be monitored in vivo. Striatal infusion of the CB1R agonist WIN55,212-2 (100 microM or 1 mM), dose-dependently decreased basal and forskolin stimulated extracellular cAMP. These effects were reversed by co-infusion of the CB1R antagonist SR141716A (30 microM), which alone had no effect up to the highest concentration tested (300 microM). These data indicate that changes in extracellular cAMP concentrations in response to CB1R stimulation can be monitored in vivo allowing the study of cannabinoid signaling in the whole animal. PMID- 15044072 TI - Changes in synaptic ultrastructure during reactive synaptogenesis in the rat dentate gyrus. AB - Advances in stereology, combined with continuing relevance to aging, as well as recovery from disease and injury make the reexamination of reactive synaptogenesis (RS) overdue. Moreover, recent mathematical models have suggested novel aspects of morphology, such as compartmentalization, may have profound effects on synaptic transmission. Given these novel findings, their correlation with other models of synaptic plasticity, and their potential significance for behavioral function, the precise nature of these changes need to be explored through quantitative morphometry. Towards this goal, the synaptic morphology of the dentate gyrus was assessed via serial electron microscopy at 3, 6, 10, 15, and 30 days following unilateral entorhinal cortex lesions. Foremost, the results showed that degree of curvature is a plastic feature of synapses. During RS, concave synapses showed an immediate/long-lasting increase in curvature, suggesting their importance in the compensation response. Flat synapses showed unique changes in growth, having implications for development and activation following synaptogenesis. Moreover, changes in size and curvature showed a different dynamic depending on proximity from damage. In the directly denervated MML, synapses showed an increase in curvature proportionate to increases in size. In the neighboring IML, however, these changes were independent-increases in curvature far surpassed synaptic growth. PMID- 15044073 TI - Asymmetric responses to rotation at high frequencies in central vestibular neurons of the alert cat. AB - The horizontal rotatory vestibulo-ocular reflex (VOR) stabilizes gaze by moving the eyes at an angular velocity proportional to head velocity, and can accomplish this for a broad range of frequencies and amplitudes of head motion. Rotation at 5 Hz and above may be processed differently than lower frequencies by the VOR network. We recorded discharges and calculated spike densities of a small sample of vestibular neurons in alert cats during low-velocity rotation at frequencies up to 8 Hz. At high frequencies, we found both vestibular-only (V-only) and eye movement-sensitive (EM) cells that generated asymmetric output signals. Asymmetry was primarily of the cutoff type, i.e., changes in spike density were smallest for rotation in the inhibitory direction. Most cells were identified as secondary neurons. The mean spike density was 23 sp/s, which was lower than previously reported in vestibular neurons of monkeys. A few neurons had very high sensitivities, associated with phase-locking, to rotation at high frequencies. In general, vestibular neurons carried a high-pass-filtered version of rotational signals. When synaptic inputs from the vestibular commissure were quantified, we found that the immediate change in probability of firing due to commissural vestibular input was inversely correlated with the degree of high-pass filtering. At high frequencies, increased asymmetry and phase-locking occurred in some neurons. A small number of neurons responded with increased probability of firing to both directions of rotation. Together, these observations suggest that high frequencies of rotation may be encoded differently than low frequencies by central vestibular neurons in alert animals. PMID- 15044074 TI - Tonic and phasic alteration in amygdala 5-HT, glutamate and GABA transmission after prefrontal cortex damage in rats. AB - The relationship between the ventromedial prefrontal cortex and the amygdala during the presentation of an unconditioned fear stimulus was assessed. Rats underwent bilateral ibotenic acid or vehicle administration into the ventromedial prefrontal cortex. Five weeks later, the behavior as well as the neurochemical changes in the amygdala was evaluated before and after a brief cat presentation. Lesioned animal freezing behavior increased 10 times when compared to controls. In the right basolateral amygdala, basal concentrations of 5-HT, 5-HIAA, glutamate and serine were elevated but basal level of GABA was diminished in lesioned animals relative to controls. Sham but not lesioned animals increased 5 HT and decreased GABA and serine levels after cat presentation. Phasic changes in glutamate were not detected either in lesioned or shams but the difference in amygdala glutamate between lesioned and shams persisted after cat presentation. These data show that increased serotonin and glutamate tone and decreased gabaergic tone in the amygdala correlate to elevated fear and anxiety after prefrontal cortex ibotenic acid lesion. The lesion also seems to produce a failure of adaptive changes in neurotransmitter systems revealing lost of control of the ventromedial prefrontal cortex over the amygdala in frightening situations. PMID- 15044075 TI - Ovarian steroid hormones modulate circadian rhythms of neuroendocrine dopaminergic neuronal activity. AB - Dopamine (DA) is the primary inhibitor of prolactin (PRL) secretion. Three populations of neuroendocrine dopaminergic neurons (NDNs) designated tuberoinfundibular (TIDA), tuberohypophyseal (THDA) and periventricular hypophyseal DAergic (PHDA) neurons regulate PRL secretion. Given that ovarian steroids modulate both DA release and PRL secretion independently, we characterized the role of steroid hormones in coupling rhythmic NDN activity and PRL secretion. OVX rats under a standard 12:12 L:D cycle (L:D), constant dark (DD), or a 6-h phase-delayed L:D cycle (pdL:D) were treated with Estradiol-17beta (E) or E and Progesterone (E+P). NDN activity, defined by DA:DOPAC ratio in nerve terminals, was determined by HPLC-EC. E or E+P stimulated PRL surges in L:D that persisted under DD. In TIDA neurons, E or E+P treatment reduced the amount of DA released under L:D and DD and advanced the rhythm of DA turnover. E and E+P treatment reduced THDA and PHDA neuron activity under L:D, but did not affect these rhythms under DD. Circadian rhythms of PRL, corticosterone and DA turnover in NDN terminals from steroid treated rats entrained to a pdL:D cycle within 7 days. Therefore, ovarian steroids differentially adjust the timing and magnitude of NDN activity to facilitate coupling of DA release and PRL secretion. PMID- 15044076 TI - Anticonvulsant effect of GMP depends on its conversion to guanosine. AB - Studies on the purinergic system normally deal with adenine-based purines, namely, adenine nucleotides and adenosine. However, a guanine-based purinergic system may also have important neuromodulatory roles. Guanine-based purines exert trophic effects on neural cells, protect brain slices in a model of hypoxia and stimulate glutamate uptake. In vivo, both guanosine 5'-monophosphate (GMP) and guanosine (GUO) protected against seizures. In this study, we investigated if the anticonvulsant effect of GMP is mediated by guanosine and if guanosine or GMP treatments were able to increase adenosine levels. Intraperitoneal (i.p.) treatments with 7.5 mg/kg GMP or guanosine prevented 50% of seizures by quinolinic acid (QA) and increased guanosine cerebrospinal fluid (CSF) levels around twofold and threefold, respectively; GMP and adenosine levels remained unchanged. Intracerebroventricular treatment with 960 nmol GMP prevented 80% of seizures and the 5'-nucleotidase inhibitor alpha-beta-methyleneadenosine 5' diphosphate (AOPCP), when injected 3 min before, reduced this anticonvulsant effect to 30% protection as well as significantly decreased the conversion of GMP into guanosine measured in the CSF. This study shows that the previously reported effect of GMP as an anticonvulsant seems to be related to its ability to generate guanosine through the action of ecto-5'-nucleotidase. PMID- 15044077 TI - The effect of acute stress on LTP and LTD induction in the hippocampal CA1 region of anesthetized rats at three different ages. AB - Not all experiences are memorized equally well. Especially, some types of stress are unavoidable in daily life and the stress experience can be memorized for life. Previous evidence has showed that synaptic plasticity, such as long-term potentiation (LTP) that may be the major cellular model of the mechanism underlying learning and memory, is influenced by behavioral stress. However, the effect of behavioral stress on age-related synaptic plasticity in vivo was primarily known. Here we found that the LTP induction in the hippocampal CA1 region of anesthetized rats obviously showed inverted-U shape related to ages (4, 10 and 74 weeks old rats), but low-frequency stimulation was unable to induce reliable long-term depression (LTD) in these animals. Furthermore, acute elevated platform (EP) stress enabled reliable LTD significantly and completely blocked LTP induction at these ages. Importantly, LTD after exposure to acute EP stress showed similar magnitude over these ages. The present results that stress enables LTD but impairs LTP induction at these three ages strengthen a view that stress experience-dependent LTD (SLTD) may underlie stress form of aberrant memories. PMID- 15044078 TI - Circadian expression of clock gene in the optic tectum of Japanese quail. AB - The physiological activity of avian optic tectum (TeO) is known to be regulated by the circadian system. In a previous study, we found clock gene expression in the TeO of Japanese quail. Here we report rhythmic expression of the Per2 gene in the stratum griseum et fibrosum (SGF) of the TeO under a light--dark (LD) cycle, constant darkness (DD), and constant light (LL) conditions. However, light pulse did not affect Per2 expression in the TeO. These results suggest that light stimulus and melatonin rhythm are not essential for rhythmic expression of Per2 in the avian TeO in spite of the localization of melatonin receptors and retinal input. PMID- 15044079 TI - Oxygen sensing and oxidant/redox-related pathways. AB - What is the nature of the oxygen sensor(s) and how do organisms sense variations in oxygen? A progressive rise of oxidative stress due to the altered reduction oxidation (redox) homeostasis appears to be one of the hallmarks of the processes that regulate gene transcription. Dynamic changes in oxygen homeostasis and its close association with redox equilibrium, therefore, constitute a signaling mechanism for the expression/activation of oxygenes. This variation subsequently regulates the compartmentalization and functioning of HIF-1alpha and NF-kappaB. In addition, oxygen-evoked regulation of HIF-1alpha and NF-kappaB is closely coupled with intracellular redox state, such that modulating redox equilibrium affects their responsiveness at the molecular level (expression/transactivation). Interestingly, are these particular transcription factors potential oxygen sensors? The basic components of the intracellular oxidative/redox machinery and its crucial regulation of oxygen- and redox-sensitive transcription factors may help understand the network of oxygen sensing mechanisms and redox-related pathways. PMID- 15044080 TI - Escherichia coli mRNAs with strong Shine/Dalgarno sequences also contain 5' end sequences complementary to domain # 17 on the 16S ribosomal RNA. AB - A well-established feature of the translation initiation region, which attracts the ribosomes to the prokaryotic mRNAs, is a purine rich area called Shine/Dalgarno sequence (SD). There are examples of various other sequences, which despite having no similarity to an SD sequence are capable of enhancing and/or initiating translation. The mechanisms by which these sequences affect translation remain unclear, but a base pairing between mRNA and 16S ribosomal RNA (rRNA) is proposed to be the likely mechanism. In this study, using a computational approach, we identified a non-SD signal found specifically in the translation initiation regions of Escherichia coli mRNAs, which contain super strong SD sequences. Nine of the 11 E. coli translation initiation regions, which were previously identified for having super strong SD sequences, also contained six or more nucleotides complementary to box-17 on the 16S rRNA (nucleotides 418 554). Mutational analyses of those initiation sequences indicated that when complementarity to box-17 was eliminated, the efficiency of the examined sequences to mediate the translation of chloramphenicol acetyltransferase (CAT) mRNA was reduced. The results suggest that mRNA sequences with complementarity to box-17 of 16S rRNA may function as enhancers for translation in E. coli. PMID- 15044081 TI - Hepatocyte growth factor/scatter factor is not a potent regulator of anabolic and catabolic gene expression in adult human articular chondrocytes. AB - Objective. Hepatocyte growth factor (HGF) has been reported to be present in articular cartilage and to be a potentially important inducing factor of anabolic and catabolic activity in chondrocytes. The aim of this study was to determine the expression levels of full length-functional-hgf and its receptor c-met in normal and osteoarthritic cartilage and the effect of HGF on anabolic and catabolic gene expression in adult human articular chondrocytes. Methods. Isolated adult human articular chondrocytes were stimulated for 48h with HGF (1, 10, and 100ng/ml). Synthesis of proteoglycans was determined by [(35)S]sulfate incorporation. mRNA levels for anabolic and catabolic genes as well as c-met and (functional) hgf were quantified using real-time PCR. Additionally, in situ mRNA expression levels of hgf and c-met were quantitatively measured from RNA directly isolated from normal and osteoarthritic adult human articular cartilage. Results. Proteoglycan synthesis in adult human articular chondrocytes was not stimulated by HGF nor was a selection of catabolic genes (collagenases and aggrecanases). Normal adult articular chondrocytes expressed only very low levels of hgf mRNA. Slightly higher levels of hgf were detected in chondrocytes isolated from osteoarthritic cartilage. Significant c-met expression was detected in both sample types. Conclusion. Despite the expression of its receptor c-met and its presence in articular cartilage, HGF does not appear to be a potent player in cartilage matrix turnover, at least not in terms of anabolic and catabolic gene expression in normal adult articular cartilage. PMID- 15044082 TI - High-level expression of a codon optimized recombinant dust mite allergen, Blo t 5, in Chinese hamster ovary cells. AB - Blo t 5 is a major allergen from house dust mite Blomia tropicalis. Purification of native Blo t 5 (nBlo t 5) from whole dust mite extract is tedious and gave low yield. In this study, we demonstrated that codon optimization facilitated high level expression of Blo t 5 in Chinese hamster ovary (CHO)-K1 cells and thus allows production of sufficient recombinant cBlo t 5 for specific immunotherapy. A codon optimized Blo t 5 gene was synthesized by PCR and the codon optimized or wild-type Blo t 5 gene in pcDNA3.0 was transfected into CHO-K1 cells and stably selected with Geneticin (G418). Western-immunoblot analysis of spent culture media detected a positive band at 14kDa for the codon optimized but not wild-type gene transfectants. In addition, a stable CHO-K1 clone produced up to 13 mg/L of the cBlo t 5 protein having a high correlation of human IgE reactivities and allergenicity to the native Blo t 5, thus indicating proper conformation of this recombinant form. PMID- 15044083 TI - Integrin-linked kinase function is required for transforming growth factor beta mediated epithelial to mesenchymal transition. AB - The role of integrin-linked kinase (ILK) in transforming growth factor beta (TGFbeta)-mediated epithelial to mesenchymal transition was investigated. A stable transfection of dominant-negative ILK results in the prevention of TGFbeta mediated E-cadherin delocalization. TGFbeta-mediated phosphorylation of Akt at Ser-473 was inhibited by dominant-negative ILK and PI3K inhibitors, LY294002 and wortmannin. Treatment with TGFbeta stimulated induction of Akt and ILK kinase activity in HaCat control cells. This increased ILK activity by TGFbeta was lowered by PI3K inhibitor, LY294002. In addition, PI3K inhibitor, dominant negative Akt, and dominant-negative ILK could not block TGFbeta-mediated C terminal phosphorylation of Smad2. Taken together, these data suggest that PI3K ILK-Akt pathway that is independent of the TGFbeta-induced Smad pathway is required for TGFbeta-mediated epithelial to mesenchymal transition. PMID- 15044084 TI - Cisplatin resistance is associated with deregulation in protein kinase C-delta. AB - Proteolytic activation of protein kinase C (PKC)-delta has been associated with cell death induced by the DNA damaging agent cisplatin. In the present study, we have examined if PKCdelta is affected when cells acquire resistance to cisplatin. The level of PKCdelta was elevated in cisplatin-resistant HeLa (HeLa/CP) cells compared to parental HeLa cells. Prolonged cellular exposure to the PKC activator phorbol-12,13-dibutyrate (PDBu), caused downregulation of PKCdelta in HeLa cells but not in HeLa/CP cells. Treatment of HeLa cells with PDBu resulted in the translocation of PKCdelta from the cytosol to the membrane but it failed to induce PKCdelta translocation in HeLa/CP cells. PDBu, however, induced translocation and downregulation of PKCalpha in both HeLa and HeLa/CP cells. The ability of PDBu to enhance cisplatin-induced cell death was attenuated in cisplatin-resistant HeLa cells. Thus, a deregulation in PKCdelta was associated with reduced cellular sensitivity to cisplatin. PMID- 15044085 TI - Effects of prostaglandin D2 on helper T cell functions. AB - Prostaglandin (PG) D(2) is abundantly produced by mast cells in the sites of allergic inflammations and acts on various cell types through its receptors DP and CRTH2. Among human T cells, CRTH2 is preferentially expressed on Th2-type cells. However, distribution of DP among T cells and impacts of CRTH2- and DP mediated signals on T cell functions are presently unclear. Here, we show that CD4(+) and CD8(+) T cells producing IFN-gamma and IL-2 were reduced by DP mediated signals, while CRTH2-mediated signals enhanced IL-2, IL-4, IL-5, and IL 13 production by Th2 cells. CRTH2 signals also caused up-regulation of CD11b and CD40L in resting Th2 cells. RT-PCR analysis revealed distribution of DP among Th cell subsets. On CRTH2(+) Th2 cells, the CRTH2-mediated PGD(2) effects were dominantly observed. Thus, PGD(2) favors Th2 functions through CRTH2 while restraining Th1 functions via DP, which may contribute to development of Th2 dominated status in allergic inflammations. PMID- 15044086 TI - Antigenic activation of Th1 cells in the gastric mucosa enhances dysregulated apoptosis and turnover of the epithelial cells. AB - Colonization of Helicobacter pylori in the stomach leads to chronic gastritis with massive infiltration by Th1 cells. To assess a role played by those T cells in the remodeling of gastric epithelium, we activated gastric T cells utilizing mice with CD4 T cells bearing transgenic TCR with or without deficiency in either IL-4 or IFN-gamma or IL-12. Mice developed gastritis upon injection of an antigen into gastric mucosa. While neutrophil infiltration occurred even with a control antigen, infiltration by transgenic T cells was dependent on the specific antigen. The numbers of epithelial cells undergoing apoptosis and regeneration were increased in the mice with infiltrating T cells producing IFN-gamma and the alignment of those cells in the glands was markedly dysregulated. In contrast, mice deficient in Th1 response showed no increase in cell division and apoptosis of epithelial cells. Thus, Th1 type T cells infiltrating into gastric mucosa play an independent role in controlling turnover of epithelial cells. PMID- 15044087 TI - The IL-1 receptor accessory protein is essential for PI 3-kinase recruitment and activation. AB - Interleukin-1 (IL-1) binds to its type I receptors (IL-1R), which in complex with IL-1R accessory protein (IL-1R AcP) induces various intracellular signaling events. We report here that IL-1 triggers the recruitment of phosphoinositide 3 kinase (PI 3-kinase) to a signaling complex and induces its lipid kinase activity in a biphasic manner. This IL-1-induced complex consists of IL-1R, IL-1R AcP, PI 3-kinase, and the IL-1-receptor-associated kinase (IRAK). Deletion of the C terminus 27 amino acids of IL-1R AcP resulted in a mutant, CDelta27, that could not recruit PI 3-kinase to the signalsome nor stimulate PI3-kinase activity. Moreover, CDelta27 functioned as a dominant-negative mutant that inhibited IL-1 induced PI 3-kinase and NFkappaB activation. CDelta27, however, had no effect on IL-1-dependent activation of the Jun N-terminal kinase (JNK), indicating that distinct regions of IL-1R AcP mediate the activation of PI 3-kinase and JNK. Thus, our results identified a functional region in the IL-1R AcP required for the recruitment and activation of PI 3-kinase. PMID- 15044088 TI - An observation of non-superimposable stereogeometrical features in a non-chiral one-component beta-Ala model peptide. AB - This paper describes the chemical synthesis and crystal molecular conformation of a non-chiral beta-Ala containing model peptide Boc-beta-Ala-Acc5-OCH3. The analysis revealed the existence of two crystallographically independent molecules A and B, in the asymmetric unit. Unexpectedly, while the magnitudes of the backbone torsion angles in both molecules are remarkably similar, the signs of the corresponding torsion angles are reverse therefore, inclining us to suggest the existence of non-superimposable stereogeometrical features in a non-chiral one-component beta-Ala model system. The critical mu torsion angle around CbetaH2 CalphaH2 bond of the beta-Ala residue represents a typical gauche orientation i.e., mu = 67.7 degrees in A and mu = -61.2 degrees in B, providing the molecule an overall crescent shaped topology. The observed conformation contrasts markedly to those determined for the correlated non-chiral model peptides: Boc-beta-Ala Acc6-OCH3 and Boc-beta-Ala-Aib-OCH3 signifying the role of stereocontrolling elements since the stereochemically constrained Calpha, alpha-disubstituted glycyl residues (e.g., Acc5, Acc6, and the prototype Aib) are known to strongly restrict the peptide backbone conformations in the 3(10)/alpha-helical-regions ( phi approximately +/-60+/-20 degrees, psi approximately +/-30+/-20 degrees) of the Ramachandran map. Unpredictably, the preferred, phi, psi torsion angles of the Acc5 residue fall outside the helical regions of the Ramachandran map and exhibit opposite-handed twists for A and B. The implications of the semi-extended conformation of the Acc5 residue in the construction of backbone-modified novel scaffolds and peptides of biological relevance are highlighted. Taken together, the results indicate that in short linear beta-Ala containing peptides specific structural changes can be induced by selective substitution of non-coded linear- or cyclic symmetrically Calpha,alpha-disubstituted glycines, reinstating the hypothesis that in addition to conformational restrictions, the chemical nature of the neighboring side-chain substituents and local environments collectively influences the stabilization of folding-unfolding behavior of the two methylene units of a beta-Ala residue. PMID- 15044089 TI - Molecular dynamics simulation of the C-terminal sterile alpha-motif domain of human p73alpha: evidence of a dynamical relationship between helices 3 and 5. AB - We used molecular dynamics simulation to evaluate the association properties of C terminal sterile alpha-motif (SAM) domain of human p73alpha. To test the dimerization propensity of this structure we carried out four simulations: EphB2 X-ray dimer, p73 modeled dimer, p73 NMR monomer, and p73 modeled monomer with an elongated helix 5. The results show a direct interaction between helix 5 and helix 3 since a conformational collapse of helix 3 is observed when dimer contact and/or an elongation of helix 5 is introduced by modeling in p73 SAM domain. On the basis of these results we suggest that the recognition properties of the SAM domains may be modulated by the conformational state of helix 5. PMID- 15044090 TI - Induction of endogenous glutathione by the chemoprotective agent, 3H-1,2-dithiole 3-thione, in human neuroblastoma SH-SY5Y cells affords protection against peroxynitrite-induced cytotoxicity. AB - Substantial evidence suggests that peroxynitrite generated from the bi-radical reaction of nitric oxide and superoxide is critically involved in the pathogenesis of neurodegenerative disorders, such as Parkinson's disease. Reaction with sulfhydryl (SH)-containing molecules has been proposed to be a major detoxification pathway of peroxynitrite in biological systems. This study was undertaken to determine if chemically elevated intracellular reduced glutathione (GSH), a major SH-containing biomolecule, affords protection against peroxynitrite-mediated toxicity in cultured neuronal cells. Incubation of human neuroblastoma SH-SY5Y cells with the unique chemoprotectant, 3H-1,2-dithiole-3 thione (D3T), led to a significant elevation of cellular GSH in a concentration dependent fashion. To examine the protective effects of D3T-induced GSH on peroxynitrite-mediated toxicity, SH-SY5Y cells were pretreated with D3T and then exposed to either the peroxynitrite generator, 3-morpholinosydnonimine (SIN-1), or the authentic peroxynitrite. We observed that D3T-pretreated cells showed a markedly increased resistance to SIN-1- or authentic peroxynitrite-induced cytotoxicity, as assessed by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium reduction assay. Conversely, depletion of cellular GSH by buthionine sulfoximine (BSO) caused a marked potentiation of SIN-1- or authentic peroxynitrite-mediated cytotoxicity. To further demonstrate the causal role for GSH induction in D3T mediated cytoprotection, SH-SY5Y cells were co-treated with BSO to abolish D3T induced GSH elevation. Co-treatment of the cells with BSO was found to significantly reverse the protective effects of D3T on SIN-1- or authentic peroxynitrite-elicited cytotoxicity. Taken together, this study demonstrates for the first time that D3T can induce GSH in cultured SH-SY5Y cells, and that the D3T-augmented cellular GSH defense affords a marked protection against peroxynitrite-induced toxicity in cultured human neuronal cells. PMID- 15044091 TI - An algorithm for selection of functional siRNA sequences. AB - Randomly designed siRNA targeting different positions within the same mRNA display widely differing activities. We have performed a statistical analysis of 46 siRNA, identifying various features of the 19bp duplex that correlate significantly with functionality at the 70% knockdown level and verified these results against an independent data set of 34 siRNA recently reported by others. Features that consistently correlated positively with functionality across the two data sets included an asymmetry in the stability of the duplex ends (measured as the A/U differential of the three terminal basepairs at either end of the duplex) and the motifs S1, A6, and W19. The presence of the motifs U1 or G19 was associated with lack of functionality. A selection algorithm based on these findings strongly differentiated between the two functional groups of siRNA in both data sets and proved highly effective when used to design siRNA targeting new endogenous human genes. PMID- 15044092 TI - Binding crevice for TT-232 in a homology model of type 1 somatostatin receptor. AB - Somatostatin receptor type 1 was modelled based on the atomic structure of bovine rhodopsin. Possible ways of binding interaction between somatostatin receptor type 1 and TT-232, a cycloheptapeptide analogue of somatostatin with broad therapeutic potential, were analysed by molecular docking. The twelve TT-232 conformations, obtained by NMR measurements in H(2)O-D(2)O mixture, were similar, disclosing a consensus backbone conformation. Several residues interacting with TT-232, such as Val133, Asp137 (helix 3), Arg197 (helix 4), Phe287, Gln291, Asn294 (helix 6), Ser305, and Tyr313 (helix 7), were found. In accordance, in vitro binding experiments indicated high-affinity binding of TT-232 to (125)I labelled somatostatin sites in brain membranes. The single binding crevice obtained by docking may allow the design and discovery of new peptidomimetics of TT-232 in the future. PMID- 15044093 TI - Curcumin blocks NF-kappaB and the motogenic response in Helicobacter pylori infected epithelial cells. AB - Infection of epithelial cells by the microbial pathogen Helicobacter pylori leads to activation of the transcription factor nuclear factor kappaB (NF-kappaB), the induction of pro-inflammatory cytokine/chemokine genes, and the motogenic response (cell scattering). Here we report that H. pylori-induced NF-kappaB activation and the subsequent release of interleukin 8 (IL-8) are inhibited by curcumin (diferuloylmethane), a yellow pigment in turmeric (Curcuma longa L.). Our results demonstrate that curcumin inhibits IkappaBalpha degradation, the activity of IkappaB kinases alpha and beta (IKKalpha and beta), and NF-kappaB DNA binding. The mitogen-activated protein kinases (MAPK), extracellular signal regulated kinases 1/2 (ERK1/2) and p38, which are also activated by H. pylori infection, were not inhibited by curcumin. Further, the H. pylori-induced motogenic response was blocked by curcumin. We conclude that curcumin, due to inhibition of NF-kappaB activation and cell scattering, should be considered as a potential therapeutic agent effective against pathogenic processes initiated by H. pylori infection. PMID- 15044094 TI - Transgenic mice overexpressing cyclophilin A are resistant to cyclosporin A induced nephrotoxicity via peptidyl-prolyl cis-trans isomerase activity. AB - Cyclosporin A (CsA) suppresses immune reaction by inhibiting calcineurin activity after forming complex with cyclophilins and is currently widely used as an immunosuppressive drug. Cyclophilin A (CypA) is the most abundantly and ubiquitously expressed family member of cyclophilins. We previously showed that CsA toxicity is mediated by ROS generation as well as by inhibition of peptidyl prolyl cis-trans isomerase (PPIase) activity of CypA in CsA-treated myoblasts [FASEB J. 16 (2002) 1633]. Since CsA-induced nephrotoxicity is the most significant adverse effect in its clinical utilization, we here investigated the role of CsA inhibition of CypA PPIase activity in its nephrotoxicity using transgenic mouse models. Transgenic mice of either wild type (CypA/wt) or R55A PPIase mutant type (CypA/R55A), a dominant negative mutant of CypA PPIase activity, showed normal growth without any apparent abnormalities. However, CsA induced nephrotoxicity was virtually suppressed in CypA/wt mice, but exacerbated in CypA/R55A mice, compared to that of littermates. Also, life expectancy was extended in CypA/wt mice and shortened in CypA/R55A mice during CsA administration. Besides, CsA-induced nephrotoxicity was inversely related to the levels of catalase expression and activity. In conclusion, our data provide in vivo evidence that supplement of CypA PPIase activity allows animal's resistance toward CsA-induced nephrotoxicity. PMID- 15044095 TI - Aromatic N-hydroxyguanidines as new reduction cosubstrates for dopamine beta hydroxylase. AB - Conversion of neurotransmitter dopamine into norepinephrine is catalyzed by dopamine beta-hydroxylase (DbH). The reaction requires the presence of both molecular oxygen and a reducing cosubstrate, the assumed physiological cosubstrate being ascorbic acid. We have investigated the ability of a new family of molecules, N-aryl-N'-hydroxyguanidines, to serve as cosubstrates for DbH. N-(4 Methoxyphenyl)-N'-hydroxyguanidine proved to be an efficient reducing agent for DbH. The complete N-hydroxyguanidine moiety was required for activity, as any modification of this function resulted in non-cosubstrate compounds. Moreover, analysis of the products formed from N-(4-methoxyphenyl)-N'-hydroxyguanidine showed that the main oxidation product was a nitrosoimine. Modification of the aromatic para-substituent evidenced an influence of its electronic properties on the catalytic activity whereas steric factors seemed less important. In addition, changing the methoxy-substituent from the para- to the ortho-position led to an inactive compound. Our results demonstrate that N-aryl-N'-hydroxyguanidines are new efficient reducing cosubstrates for DbH and prove that specific interactions with the reducing cosubstrate do take place at the active site of the enzyme. PMID- 15044096 TI - Effect of exogenous E2F-1 on the expression of common chromosome fragile site genes, FHIT and WWOX. AB - The expression of two tumor suppressor genes, fragile histidine triad (FHIT) and WW domain containing oxidoreductase (WWOX), encompassing common chromosome fragile regions, FRA3B at 3p14.2 and FRA16D at 16q23, is altered in many epithelial tumors. Since DNA sequence search shows that the FHIT gene has the E2F 1 recognition site in 5'] region, which regulates cell cycle, we tested the effect of E2F-1 overexpression in tumor cells. Ectopic E2F-1 expression led to an increase of Fhit and Wwox expression in allele remaining tumor cells and resulted in induction of apoptosis. Reporter assay showed that the E2F-1 site in FHIT 5' region was involved in the down-stream transcription after exogenous E2F-1 introduction. Chromatin immunoprecipitation detected exogenous E2F-1 binding to the recognition site in FHIT 5' region. The data suggest that E2F-1 overexpression plays a role in suppression of tumor, at least in part trough transcriptional regulation of FHIT and relevant activation of WWOX. PMID- 15044097 TI - In vitro transdifferentiation of adult pancreatic acinar cells into insulin expressing cells. AB - Despite a recent breakthrough in human islet transplantation for treating diabetes mellitus, the limited availability of insulin-producing tissue is still a major obstacle. Here, we studied whether adult pancreatic acinar cells have the potential to transdifferentiate into islet or beta cells. Pancreatic acini were isolated from 7- to 8-weeks-old male Sprague-Dawley rats and cultured in suspension. Within 1 week, most of the acinar cells lost amylase expression and converted to cells with a duct cell phenotype. Insulin-positive cells were also observed, mainly at the periphery of the acini-derived spheroids. Insulin gene and protein expression was increased. Presence of a few insulin-positive cells coexpressing cytokeratins suggests that a spontaneous acinar to ductal cell transdifferentiation process was further going on towards beta cells. This study provides the first evidence that adult pancreatic acinar cells could be differentiated into insulin-expressing cells in vitro. PMID- 15044098 TI - Structure-function studies of the Vitreoscilla hemoglobin D-region. AB - The D-region connecting helices C and E of Vitreoscilla hemoglobin (VHb) appears disordered in the crystal structure. Six site-directed mutants in this region were made to investigate its possible functions. The mutant VHb's were analyzed using UV-visible and FTIR spectroscopy, using primarily the CO liganded forms, and their heme/protein ratios were determined. The results implicate Asp44, Arg47, and Glu49 as especially important in heme-globin interactions and ligand binding, and enabled construction of a model in which the D-region forms a loop that protrudes upward over the heme. Interactions between VHb (wild type and the D-region mutants) with the flavin domain of 2,4-DNT dioxygenase from Burkholderia were tested using bacterial two-hybrid screening. There was a correlation between the extent of the D-loop perturbation predicted for each mutant and the amount of the reduction in VHb-flavin domain interaction, suggesting that this region may be more generally involved in binding of VHb to flavoproteins. PMID- 15044099 TI - Induction of cell death in activated hepatic stellate cells by targeted gene expression of the thymidine kinase/ganciclovir system. AB - Liver fibrosis is the result from a relative imbalance between synthesis and degradation of matrix proteins. Following liver injury of any etiology, hepatic stellate cells undergo a response known as activation, which is the transition of quiescent cells into proliferative, fibrogenic, and contractile myofibroblasts. Upon this cellular transdifferentiation the effector cell becomes the major source of fibrillar and non-fibrillar matrix proteins resulting in excessive scar formation and cirrhosis, the end stage of fibrosis. Concomitant with progressive liver fibrosis, the tissue inhibitor of metalloproteinases-1 (TIMP-1) is strongly activated in hepatic stellate cells. We have developed a recombinant replication defective adenovirus in which the TIMP-1 promoter is coupled to the herpes simplex virus thymidine kinase gene rendering activated hepatic stellate cells susceptible to ganciclovir. This novel targeted suicide gene approach was validated in a culture model considered to reflect an accelerated time course of the cellular and molecular events that occur during liver fibrosis. We demonstrate that transfer of the suicide gene to culture-activated hepatic stellate cells results in a strong expression of the respective transgene as assessed by Northern blot and Western blot analyses. The enzyme catalyzed the proper conversion of its prodrug subsequently initiating programmed cell death as estimated by caspase-3 assay and Annexin V-Fluos staining. Altogether, these results indicate that induction of programmed cell death is a promising approach to eliminate fibrogenic HSC. PMID- 15044100 TI - Human MCRS2, a cell-cycle-dependent protein, associates with LPTS/PinX1 and reduces the telomere length. AB - Human LPTS/PinX1 is a telomerase-inhibitory protein, which binds to the telomere protein Pin2/TRF1 and the catalytic subunit hTERT of telomerase. To explore the proteins that might be involved in the telomerase pathway, we performed a yeast two-hybrid screening with LPTS/PinX1 as the bait. A novel gene, MCRS2, encoding for an isoform of MCRS1/p78 and MSP58 was isolated. The expression of MCRS2 protein is cell-cycle dependent, accumulating in the very early S phase. MCRS2 interacts with LPTS/PinX1 in vitro, in vivo and colocalizes with LPTS/PinX1 in cells. MCRS2 and its amino terminus inhibit telomerase activity in vitro and long term overexpression of MCRS2 in SMMC-7721 cells results in a gradual and progressive shortening of telomeres. Our findings suggest that MCRS2 might be a linker between telomere maintenance and cell-cycle regulation. PMID- 15044101 TI - Tertiary structural changes associated with iron binding and release in hen serum transferrin: a crystallographic and spectroscopic study. AB - The iron binding and release of serum transferrin are pH-dependent and accompanied by a conformational change between the iron-bound (holo-) and iron free (apo-) forms. We have determined the crystal structure of apo-hen serum transferrin (hAST) at 3.5A resolution, which is the first reported structure to date of any full molecule of an apo-serum transferrin and studied its pH dependent iron release by UV-vis absorption and near UV-CD spectroscopy. The crystal structure of hAST shows that both the lobes adopt an open conformation and the relative orientations of the domains are different from those of apo human serum transferrin and human apolactoferrin but similar to that of hen apo ovotransferrin. Spectroscopic analysis reveals that in hen serum transferrin, release of the first iron starts at a pH approximately 6.5 and continues over a broad pH range (6.5-5.2). The complete release of the iron, however, occurs at pH approximately 4.0. The near UV-CD spectra show alterations in the microenvironment of the aromatic residues surrounding the iron-binding sites. PMID- 15044102 TI - Human hepatic cell uptake of resveratrol: involvement of both passive diffusion and carrier-mediated process. AB - This work reports significant advances on the transport in hepatic cells of resveratrol, a natural polyphenol with potent protective properties. First, we describe a new simple technique to qualitatively follow resveratrol cell uptake and intracellular distribution, based on resveratrol fluorescent properties. Second, the time-course study and the quantification of (3)H-labelled resveratrol uptake have been performed using human hepatic derived cells (HepG2 tumor cells) and hepatocytes. The temperature-dependence of the kinetics of uptake as well as the cis-inhibition experiments agree with the involvement of a carrier-mediated transport in addition to passive diffusion. The decrease of passive uptake resulting from resveratrol binding to serum proteins brings to light a mediated mechanism in physiological situation. PMID- 15044103 TI - Quinone analogues regulate mitochondrial substrate competitive oxidation. AB - Quinone derivatives are among the rare compounds successfully used as therapeutic reagents to fight mitochondrial diseases. However, their beneficial effect appears to depend on their side chain which presumably governs their interaction with the respiratory chain. The effect of four quinone derivatives was comparatively studied on NADH- and succinate-competitive oxidation by a sub mitochondrial fraction. Under our experimental conditions, the less hydrophobic derivatives (menadione, duroquinone) poorly affected electron flow from either NADH or succinate to oxygen, yet readily diverting electrons from isolated complex I. This latter effect was abolished by succinate addition. More hydrophobic derivatives (idebenone, decylubiquinone) stimulated oxygen uptake from succinate. But while NADH oxidation was slightly inhibited by idebenone, it was somewhat increased by decylubiquinone. As a result, idebenone strongly favoured succinate over NADH oxidation. This study therefore suggests that any therapeutic use of quinone analogues should take into account their specific effect on each respiratory chain dehydrogenase. PMID- 15044104 TI - Effect of glycine substitutions on alpha5(IV) chain structure and structure phenotype correlations in Alport syndrome. AB - The phenotype variety caused by glycine substitutions in alpha5(IV) chain in X linked Alport syndrome (XLAS) prompted the complexity of structure changes of alpha5(IV) chain that was little to know now. In this study, we expressed a domain of alpha5(IV) chain containing different glycine substitutions (G1015V and G1030S, respectively) which were revealed in two XLAS pedigrees with different phenotype severities and the corresponding domain of a control in Escherichia coli. The recombinant proteins were characterized by immunoblot and mass spectrometry and analyzed the secondary structure by using circular dichroism (CD) spectroscopy. CD analysis showed that the recombinant protein containing G1015V mutation identified in the pedigree of juvenile-onset XLAS exhibited 12.9% alpha-helix that was not found in the control recombinant protein. The spectrum of the recombinant protein containing G1030S mutation identified in the pedigree of adult-onset XLAS was slightly different from that of the control, that is, mostly with the random coil and the beta-sheet, while without alpha-helix. These results demonstrated that two kinds of glycine substitutions, although in the same domain of alpha5(IV) chain, displayed the distinctly different secondary structures. The changes of the secondary structure could explain the phenotypic diversities of XLAS, which would be hardly understood solely by analyzing genomic DNA or mRNA of alpha5(IV) chain. PMID- 15044105 TI - Denaturation of dsDNA by p53: fluorescence correlation spectroscopy study. AB - p53 activates transcription through interaction with specific DNA sequences in gene promoters. It also regulates DNA replication, recombination, and repair apparently through interactions with DNA intermediates of these reactions. Biochemical activities relevant for these functions of p53 include binding to the ends and internal segments of single-stranded DNA molecules, catalysis of DNA renaturation, and strand exchange. We report a novel activity of p53, its ability to denature double-stranded DNA molecules aggregated by basic peptides. Stable complexes of coiled single-stranded DNA molecules with basic peptides are formed in this reaction. Thus, complementary to the ability to catalyze DNA renaturation, p53 denatures double-stranded DNA when the latter reaction is thermodynamically favorable. This p53 activity, along with its ability to interact physically with DNA helicases, may be relevant for resolving double stranded DNA intermediates and inhibition of DNA recombination, which is critical for guarding of the genome. PMID- 15044106 TI - Expression and methylation status of 14-3-3 sigma gene can characterize the different histological features of ovarian cancer. AB - We hypothesize that 14-3-3 sigma gene expression and its regulation by methylation can characterize histological types of primary human epithelial ovarian cancer. To test this hypothesis, ovarian cancer cell lines and 54 ovarian cancer tissue samples were analyzed for expression and methylation of 14-3-3 sigma gene using methylation specific PCR. The results of our experiments demonstrate that 14-3-3 sigma gene was methylated and inactivated in ES-2 ovarian cell line, which was derived from clear cell adenocarcinoma. Treatment of this cell line with demethylating agent 5-aza-2'-deoxycytidine restored the expression of 14-3-3 sigma gene. In human ovarian cancer tissues, the expression of 14-3-3 sigma protein was inactivated in most of the ovarian clear cell carcinoma tissues. Interestingly, 14-3-3 sigma protein expression was positive in significantly higher percentages of serous (89.5%), endometrioid (90%), and mucinous (81.8%) ovarian adenocarcinoma tissues. The ovarian clear cell carcinoma samples with inactivated 14-3-3 sigma protein were highly methylated, suggesting that inactivation of 14-3-3 sigma gene is through DNA methylation. Using direct DNA sequencing, 14-3-3 sigma gene methylation on all the 17 CpG sites was significantly higher in ovarian clear cell carcinoma as compared to other histological types of ovarian cancer (serous, endometrioid, and mucinous). This is the first report suggesting that 14-3-3 sigma gene expression and methylation status can characterize histological features of different types of ovarian cancer. PMID- 15044107 TI - Truncated hemoglobin o of Mycobacterium tuberculosis: the oligomeric state change and the interaction with membrane components. AB - Being an obligate aerobe, the Mycobacterium tuberculosis cells would have to evolve a mechanism to collect and deliver the hardly available O(2) to survive in granulomas and to maintain the low level of respiration during latency. The M. tuberculosis truncated hemoglobin o (trHbO), when heterologously expressed in Escherichia coli cells, was found to significantly enhance the cellular respiration and cell growth. This study was undertaken in an attempt to understand the molecular details for trHbO to promote the cellular respiration, focusing on the ways through which trHbO is recruited to the cell membrane and O(2) molecules are delivered. Our data demonstrate that the trHbO protein is able to promote the growth of E. coli cells in a fashion that depends on the presence of the respiratory chain terminal oxidase cytochrome o complex (or Cyo complex). The trHbO protein appears to interact with the Cyo B subunit of the Cyo complex directly, likely in a dynamic manner. The trHbO is also able to bind membrane lipids in a non-specific way, during the process electrostatic and hydrophobic interactions both likely exist. Besides, binding with membrane induces the dissociation of trHbO from dimers to monomers. In light of these observations, a hypothesis was made to explain how trHbO might serve as an O(2) collector and/or reservoir for M. tuberculosis cells under O(2)-limiting or lacking conditions. PMID- 15044108 TI - Multidrug-resistant tumor cells with decreased malignancy: a role for integrin alphavbeta3. AB - We studied whether acquisition of multidrug resistance (MDR) by tumor cells can alter their integrin profile and malignant behavior. Hamster fibroblast cell line HET-SR-2SC-LNM was selected for MDR, yielding the 2SC/20 subline. Compared with the parental cells, the 2SC/20 subline weakly adhered to denatured collagen (dCol) which correlated with decreased expression of alphavbeta3, a dCol receptor. Importantly, 2SC/20 subline demonstrated significantly decreased activity of collagenase MMP-2, lower ability to invade Matrigel, and attenuated metastasis in syngeneic animals. We provide evidence for the first time that selection for MDR can be associated with down-regulation of alphavbeta3 integrin, supporting our recent proof of the pro-apoptotic role of this integrin (Oncogene 20 (2001) 4710). Lack of alphavbeta3 expression may link cell survival under toxic conditions with decreased malignancy of the resulting drug resistant tumor. PMID- 15044109 TI - Arsenic trioxide uptake by human and rat aquaglyceroporins. AB - Aquaglyceroporins are channels that allow downhill movement of uncharged solutes such as glycerol and urea. Arsenic trioxide has recently been shown to be translocated by mouse mAQP7 and rat rAQP9. In this study we examined the ability of the four known human members of the aquaglyceroporin family, hAQP3, hAQP7, hAQP9, and hAQP10, to facilitate As(OH)(3) movement in Xenopus oocytes. The order of effectiveness as an As(III) transporter was found to be hAQP9 > hAQP7, with little or no transport by hAQP3 or hAQP10. From comparison with the crystal structure of the bacterial homologue GlpF and the bovine erythrocyte water channel bAQP1, AQP9 residues Phe-64 and Arg-219 are predicted to serve as part of the selectivity filter. The requirement for Phe-64 and Arg-219 in arsenic trioxide translocation was examined by site-directed mutagenesis of rAQP9, taking advantage of the fact that rat AQP9 catalyzes (73)As(OH)(3) uptake in Saccharomyces cerevisiae and in oocytes. R219A, R219K, F64A, F64T, and F64W were expressed in both yeast and oocytes, and permeability of arsenic trioxide and glycerol was measured. A lysine but not an alanine residue could substitute for the highly conserved Arg-219, indicating that a positive charge is required at the entry to the channel. In contrast, the phenylalanine residue, which is believed to position substrates near the conserved arginine, was not required for either arsenic trioxide or glycerol uptake. The results support the hypothesis that arsenic trioxide and glycerol use the same translocation pathway in AQP9. PMID- 15044110 TI - Polymorphism in the second intron of dopamine receptor D4 gene in humans and apes. AB - The dopamine receptor D4 (DRD4) has received increasing research attention in behavioral science, psychiatry, and psychopharmacology. However, the number of available genetic markers for primates is still insufficient. We identified a novel variation/polymorphism in the second intron of DRD4 in humans based on the survey of 210 Japanese: a 6bp insertion (allele frequency: 0.002) and 8bp deletion (0.024); however, 94 Hungarian Caucasians were found to be monomorphic. Polymorphisms of the homologous region were also found in a survey of 93 specimens from four species of great apes and 51 specimens from seven species of gibbons. The polymorphisms consist of both single nucleotide substitutions and variations in the number of tandem duplications of short GC-rich sequences. Because of usefulness of primates in behavioral science, this polymorphism may be a useful marker for association studies with behavioral traits in both humans and apes. PMID- 15044111 TI - The arachidonate-dependent cytoprotective signaling evoked by peroxynitrite is a general response of the monocyte/macrophage lineage. AB - U937, THP-1, and J774 cells or human monocytes and macrophages display similar levels of sensitivity to peroxynitrite and exposure to an otherwise non-toxic concentration of the oxidant in the presence of a phospholipase A(2) inhibitor was invariably associated with the onset of mitochondrial permeability transition (MPT)-dependent toxicity. These events were prevented by exogenous arachidonic acid (AA). In general, the protective concentrations of AA were greater in those cell types releasing more AA. Thus, non-toxic concentrations of peroxynitrite commit cells belonging to the monocyte/macrophage lineage to MPT-dependent toxicity that is however prevented by endogenous AA. PMID- 15044112 TI - Exogenous insulin can reverse the effects of caloric restriction on mitochondria. AB - It has been proposed that part of the anti-aging mechanism of caloric restriction (CR) involves a reduction in both the generation rate of reactive oxygen species (ROS) by mitochondria, and a reduction in peroxidizability of mitochondrial membranes. It was hypothesized that these effects may be due to upstream changes in hormonal status, since certain hormones (such as insulin) are stimulatory for ROS production, effect fatty acid composition, and are lowered by CR. To investigate this hypothesis, young male Brown-Norway rats on 55% CR (4 months duration) were subjected to insulin replacement by use of mini-osmotic pumps. ROS and free radical-induced malondialdehdye production were significantly lower in mitochondria from CR animals compared to those from fully fed, and these effects were reversed by insulin. It is concluded that the beneficial changes induced by CR, as seen at the mitochondrion, may in part be downstream effects of alterations in hormonal signalling. PMID- 15044113 TI - A bacterial expression system revisited for the recombinant production of cystine rich plant lipid transfer proteins. AB - Non-specific lipid transfer proteins (nsLTPs) are abundant and ubiquitous cystine rich proteins that are capable, in vitro, of binding lipids and hydrophobic molecules. In view to probe the lipid binding properties of the wheat nsLTP1, mutant variants may represent a powerful tool. To this end, a synthetic gene, encoding a mature wheat nsLTP1 polypeptide, was designed to ensure high level expression in Escherichia coli. The bacterial expression host strain, a translational fusion strategy, and convenient cleavage and purification procedures were optimized to produce in standard fermentation conditions, a significant amount (15 mg/L final yield), of a soluble and correctly folded recombinant nsLTP1. This highly amenable expression system is helpful in order to investigate structure-activity relationships of plant nsLTP. PMID- 15044114 TI - Gene-nutrient and gene-gene interactions of controlled folate intake by Japanese women. AB - Elevated serum total homocysteine (tHcy) levels are associated with increased risk for cardiovascular disease and dementia. The prevalence rates of homozygous mutants among Japanese women (n = 300) were 17.3%, 1.3%, 18.6%, and 5.3% for methylenetetrahydrofolate reductase (MTHFR) C677T and A1298C, reduced folate carrier (RFC-1) A80G, and methionine synthase (MS) A2756G, respectively. The tHcy value was significantly lower (p < 0.001) in young women with CC or CT of MTHFR than with TT (10.9+/-4.7 micromol/L) (n =250). Diversities of serum folate and tHcy in women with 23 combinations of different alleles at low folate intake converged to the highest (34.0+/-8.6 nmol/L) and lowest (7.6+/-1.5 micromol/L) levels, respectively, after folic acid (400 microg/day) supplementation. In the regression equation ( y= ax + b) of serum folate ( y nmol/L) plotted against mean folate intake ( x microg/day), the values of "a" were 0.032, 0.037, and 0.045 for individuals with CC, CT, and TT alleles, respectively, of MTHFR. PMID- 15044115 TI - Mapping protein matrix cavities in human cytoglobin through Xe atom binding. AB - Cytoglobin is the fourth recognized globin type, almost ubiquitously distributed in human tissues; its function is still poorly understood. Cytoglobin displays a core region of about 150 residues, structurally related to hemoglobin and myoglobin, and two extra segments, about 20 residues each, at the N- and C termini. The core region hosts a large apolar cavity, held to provide a ligand diffusion pathway to/from the heme, and/or ligand temporary docking sites. Here we report the crystal structure (2.4A resolution, R-factor 19.1%) of a human cytoglobin mutant bearing the CysB2(38) --> Ser and CysE9(83) --> Ser substitutions (CYGB*), treated under pressurized xenon. Three Xe atoms bind to the heme distal site region of CYGB* mapping the protein matrix apolar cavity. Despite the conserved globin fold, the cavity found in CYGB* is structured differently from those recognized to play a functional role in myoglobin, neuroglobin, truncated hemoglobins, and Cerebratulus lacteus mini-hemoglobin. PMID- 15044116 TI - The trouble with trans fat. PMID- 15044117 TI - Bladder cancer in women: no time to wait. AB - Bladder cancer is more common in men, but more often discovered at an advanced stage in women. Know the warning signs--and act on them quickly. PMID- 15044118 TI - Update on dietary supplements for depression. AB - When does an alternative therapy become mainstream? When enough studies show it's safe and effective. For mild to moderate depression, a handful of supplements seem headed in that direction. PMID- 15044119 TI - By the way, doctor. I've recently heard about a new cholesterol drug called Zetia. Does it work better than a statin? PMID- 15044120 TI - By the way, doctor. I was astounded to read that women who test positive for the breast cancer gene have as much as an 80% risk of getting breast cancer. Shouldn't we all be tested? PMID- 15044121 TI - Acute urinary retention: An age-old problem. PMID- 15044122 TI - Medical memo: Do physicians heal themselves? PMID- 15044123 TI - The Polypill: a simple "cure" for heart disease and stroke? PMID- 15044124 TI - On call. My wife's nutrition newsletter says that everyone over 60 should take a multivitamin with 25 micrograms of vitamin B12. We're both 60 and healthy, but the vitamin I've been taking since I read your February 2000 article on supplements has only 2.4 micrograms of B12. Am I missing out? Why are these recommendations so different? PMID- 15044125 TI - Problem gambling. AB - Addicted gamblers take bad risks with their money and their lives. They can be helped to recover both. PMID- 15044127 TI - How not to be happy. PMID- 15044126 TI - Psychosocial treatment of bipolar disorder. AB - Drug treatment is essential but insufficient. Fortunately, there are many other ways to help bipolar patients cope with their symptoms and manage their lives. PMID- 15044128 TI - Eating disorders and culture. PMID- 15044129 TI - Questions & answers. We've been told that my wife has multi-infarct dementia and that the new drug memantine may help. How is her condition different from Alzheimer's disease, and what could memantine do for her? PMID- 15044130 TI - 11 things you should talk to your doctor about. It's easy to suggest that you talk to your doctor. It's much harder to know what to say. PMID- 15044131 TI - Virtual colonoscopy: is it ready for the real world? A recent study shows that it may now be as accurate as conventional colonoscopy. PMID- 15044132 TI - Obesity in the extreme. It has risks, but surgery is the most effective treatment for people with severe obesity. PMID- 15044134 TI - Vitamin D shortfalls casting wider shadow. PMID- 15044133 TI - Snooze alarm: you need your sleep. Sleep, like diet and exercise, is an essential ingredient of good health. PMID- 15044135 TI - Stroke symptoms. PMID- 15044136 TI - Smell the coffee, feel the caffeine. PMID- 15044137 TI - By the way, doctor. I get mail extolling the virtues of chelation therapy. The claim is that it removes calcium and other metals from the walls of arteries. The key element appears to be EDTA. Since vinegar is also primarily acetic acid, I'm curious whether it could have the same effect. PMID- 15044138 TI - By the way, doctor. Is the CV Profiler a legitimate diagnostic tool for arterial elasticity? I understand that this technology just became available last year. PMID- 15044139 TI - Protecting the heart while controlling blood sugar. New guidelines help define and refine the use of two diabetes drugs that could be a boon for people who also have heart disease. PMID- 15044140 TI - Possible plaque buster offers new hope. HDL could become a treatment for heart disease. PMID- 15044141 TI - Gender bender. Awareness that heart disease affects women in different ways than men can help women get the treatments they need and pay more attention to prevention. PMID- 15044142 TI - 12 tips for safe travel with heart disease. Vacations can be great for the body and soul. Make sure your next one is good for the heart, too. PMID- 15044143 TI - When the natural pacemaker loses its groove. Here's how to recognize and treat a sick sinus. PMID- 15044144 TI - Keeping clots from mechanical heart valves. PMID- 15044145 TI - Breakthrough in treating cardiac arrest. PMID- 15044146 TI - Ask the doctor. My wife has had all of the symptoms described in a recent study showing that women have different heart attack warning signs than men. I urged her to get a stress test but she says her doctor told her that the treadmill test isn't good at diagnosing heart trouble in women. Is this the case? Thanks to the treadmill test, I avoided a heart attack. Now I want the same thing for my wife! PMID- 15044147 TI - Ask the doctor. Reports that omega-3 fats are good for the heart make me want to get more in my diet. I am allergic to fish, and my allergist told me not to take fish oil capsules. What are my options? PMID- 15044148 TI - Fiber type conversion alters inactivation of voltage-dependent sodium currents in murine C2C12 skeletal muscle cells. AB - Each skeletal muscle of the body contains a unique composition of "fast" and "slow" muscle fibers, each of which is specialized for certain challenges. This composition is not static, and the muscle fibers are capable of adapting their molecular composition by altered gene expression (i.e., fiber type conversion). Whereas changes in the expression of contractile proteins and metabolic enzymes in the course of fiber type conversion are well described, little is known about possible adaptations in the electrophysiological properties of skeletal muscle cells. Such adaptations may involve changes in the expression and/or function of ion channels. In this study, we investigated the effects of fast-to-slow fiber type conversion on currents via voltage-gated Na+ channels in the C(2)C(12) murine skeletal muscle cell line. Prolonged treatment of cells with 25 nM of the Ca2+ ionophore A-23187 caused a significant shift in myosin heavy chain isoform expression from the fast toward the slow isoform, indicating fast-to-slow fiber type conversion. Moreover, Na+ current inactivation was significantly altered. Slow inactivation less strongly inhibited the Na+ currents of fast-to-slow fiber type-converted cells. Compared with control cells, the Na+ currents of converted cells were more resistant to block by tetrodotoxin, suggesting enhanced relative expression of the cardiac Na+ channel isoform Na(v)1.5 compared with the skeletal muscle isoform Na(v)1.4. These results imply that fast-to-slow fiber type conversion of skeletal muscle cells involves functional adaptation of their electrophysiological properties. PMID- 15044149 TI - Stretch-induced nitric oxide modulates mechanical properties of skeletal muscle cells. AB - In this study, we examined the hypothesis that stretch-induced (nitric oxide) NO modulates the mechanical properties of skeletal muscles by increasing accumulation of protein levels of talin and vinculin and by inhibiting calpain induced proteolysis, thereby stabilizing the focal contacts and the cytoskeleton. Differentiating C(2)C(12) myotubes were subjected to a single 10% step stretch for 0-4 days. The apparent elastic modulus of the cells, E(app), was subsequently determined by atomic force microscopy. Static stretch led to significant increases (P < 0.01) in E(app) beginning at 2 days. These increases were correlated with increases in NO activity and neuronal NO synthase (nNOS) protein expression. Expression of talin was upregulated throughout, whereas expression of vinculin was significantly increased only on days 3 and 4. Addition of the NO donor l-arginine onto stretched cells further enhanced E(app), NOS activity, and nNOS expression, whereas the presence of the NO inhibitor N(omega)-nitro-l arginine methyl ester (l-NAME) reversed the effects of mechanical stimulation and of l-arginine. Overall, viscous dissipation, as determined by the value of hysteresis, was not significantly altered. For assessment of the role of vinculin and talin stability, cells treated with l-NAME showed a significant decrease in E(app), whereas addition of a calpain inhibitor abolished the effect. Thus our results show that NO inhibition of calpain-initiated cleavage of cytoskeleton proteins was correlated with the changes in E(app). Together, our data suggest that NO modulates the mechanical behavior of skeletal muscle cells through the combined action of increased talin and vinculin levels and a decrease in calpain mediated talin proteolysis. PMID- 15044150 TI - Changes in gene expression in gills of the euryhaline killifish Fundulus heteroclitus after abrupt salinity transfer. AB - Maintenance of ion balance requires that ionoregulatory epithelia modulate ion flux in response to internal or environmental osmotic challenges. We have explored the basis of this functional plasticity in the gills of the euryhaline killifish Fundulus heteroclitus. The expression patterns of several genes encoding ion transport proteins were quantified after transfer from near isosmotic brackish water [10 parts/thousand (ppt)] to either freshwater (FW) or seawater (SW). Many changes in response to SW transfer were transient. Increased mRNA expression occurred 1 day after transfer for Na(+)-K(+)-ATPase-alpha(1a) (3 fold), Na(+)-K(+)-2Cl(-)-cotransporter 1 (NKCC1) (3-fold), and glucocorticoid receptor (1.3-fold) and was paralleled by elevated Na(+)-K(+)-ATPase activity (2 fold). The transient increase in NKCC1 mRNA expression was followed by a later 2 fold rise in NKCC protein abundance. In contrast to the other genes studied in the present work, mRNA expression of the cystic fibrosis transmembrane conductance regulator (CFTR) Cl(-) channel generally remained elevated (2-fold) in SW. No change in protein abundance was detected, however, suggesting posttranscriptional regulation. The responses to FW transfer were quite different from those to SW transfer. In particular, FW transfer increased Na(+)-K(+)-ATPase alpha(1a) mRNA expression and Na(+)-K(+)-ATPase activity to a greater extent than did SW transfer but had no effect on V-type H(+)-ATPase expression, supporting the current suggestion that killifish gills transport Na(+) via Na(+)/H(+) exchange. These findings demonstrate unique patterns of ion transporter expression in killifish gills after salinity transfer and illustrate important mechanisms of functional plasticity in ion-transporting epithelia. PMID- 15044151 TI - TRPC4 forms store-operated Ca2+ channels in mouse mesangial cells. AB - Studies were performed to identify the molecular component responsible for store operated Ca(2+) entry in murine mesangial cells (MMC). Because the canonical transient receptor potential (TRPC) family of proteins was previously shown to comprise Ca(2+)-selective and -nonselective cation channels in a variety of cells, we screened TRPC1-TRPC7 with the use of molecular methods and the fura 2 method to determine their participation as components of the mesangial store operated Ca(2+) (SOC) channel. Using TRPC-specific primers and RT-PCR, we found that cultured MMC contained mRNA for TRPC1 and TRPC4 but not for TRPC2, TRPC3, TRPC5, TRPC6, and TRPC7. Immunocytochemical staining of MMC revealed predominantly cytoplasmic expression of TRPC1 and plasmalemmal expression of TRPC4. The role of TRPC4 in SOC was determined with TRPC4 antisense and fura 2 ratiometric measurements of intracellular Ca(2+) concentration ([Ca(2+)](i)). SOC was measured as the increase in [Ca(2+)](i) after extracellular Ca(2+) was increased from <10 nM to 1 mM in the continued presence of thapsigargin. We found that TRPC4 antisense, which reduced plasmalemmal expression of TRPC4, inhibited SOC by 83%. Incubation with scrambled TRPC4 oligonucleotides did not affect SOC. Immunohistochemical staining identified expressed TRPC4 in the glomeruli of mouse renal sections. The results of RT-PCR performed to distinguish between TRPC4 alpha and TRPC4-beta were consistent with expression of both isoforms in brain but with only TRPC4-alpha expression in MMC. These studies show that TRPC4-alpha may form the homotetrameric SOC in mouse mesangial cells. PMID- 15044153 TI - Maitotoxin-induced cell death cascade in bovine aortic endothelial cells: divalent cation specificity and selectivity. AB - The maitotoxin (MTX)-induced cell death cascade in bovine aortic endothelial cells (BAECs), a model for Ca(2+) overload-induced toxicity, reflects three sequential changes in plasmalemmal permeability. MTX initially activates Ca(2+) permeable, nonselective cation channels (CaNSC) and causes a massive increase in cytosolic free Ca(2+) concentration ([Ca(2+)](i)). This is followed by the opening of large endogenous cytolytic/oncotic pores (COP) that allow molecules <800 Da to enter the cell. The cells then lyse not by rupture of the plasmalemma but through the activation of a "death" channel that lets large proteins (e.g., 140-160 kDa) leave the cell. These changes in permeability are accompanied by the formation of membrane blebs. In this study, we took advantage of the well-known differences in affinity of various Ca(2+)-binding proteins for Ca(2+) and Sr(2+) vs. Ba(2+) to probe their involvement in each phase of the cell death cascade. Using fluorescence techniques at the cell population level (cuvette-based) and at the single-cell level (time-lapse videomicroscopy), we found that the replacement of Ca(2+) with either Sr(2+) or Ba(2+) delayed both MTX-induced activation of COP, as indicated by the uptake of ethidium bromide, and subsequent cell lysis, as indicated by the uptake of propidium iodide or the release of cell-associated green fluorescent protein. MTX-induced responses were mimicked by ionomycin and were significantly delayed in BAPTA-loaded cells. Experiments at the single-cell level revealed that Ba(2+) not only delayed the time to cell lysis but also caused desynchronization of the lytic phase. Last, membrane blebs, which were numerous and spherical in Ca(2+)-containing solutions, were poorly defined and greatly reduced in number in the presence of Ba(2+). Taken together, these results suggest that intracellular high-affinity Ca(2+)-binding proteins are involved in the MTX-induced changes in plasmalemmal permeability that are responsible for cell demise. PMID- 15044152 TI - RhoA, Rac1, and Cdc42 exert distinct effects on epithelial barrier via selective structural and biochemical modulation of junctional proteins and F-actin. AB - Epithelial intercellular junctions regulate cell-cell contact and mucosal barrier function. Both tight junctions (TJs) and adherens junctions (AJs) are regulated in part by their affiliation with the F-actin cytoskeleton. The cytoskeleton in turn is influenced by Rho family small GTPases such as RhoA, Rac1, and Cdc42, all of which constitute eukaryotic targets for several pathogenic organisms. With a tetracycline-repressible system to achieve regulated expression in Madin-Darby canine kidney (MDCK) epithelial cells, we used dominant-negative (DN) and constitutively active (CA) forms of RhoA, Rac1, and Cdc42 as tools to evaluate the precise contribution of each GTPase to epithelial structure and barrier function. All mutant GTPases induced time-dependent disruptions in epithelial gate function and distinct morphological alterations in apical and basal F-actin pools. TJ proteins occludin, ZO-1, claudin-1, claudin-2, and junctional adhesion molecule (JAM)-1 were dramatically redistributed in the presence of CA RhoA or CA Cdc42, whereas only claudins-1 and -2 were redistributed in response to CA Rac1. DN Rac1 expression also induced selective redistribution of claudins-1 and -2 in addition to JAM-1, whereas DN Cdc42 influenced only claudin-2 and DN RhoA had no effect. AJ protein localization was unaffected by any mutant GTPase, but DN Rac1 induced a reduction in E-cadherin detergent solubility. All CA GTPases increased the detergent solubility of claudins-1 and -2, but CA RhoA alone reduced claudin 2 and ZO-1 partitioning to detergent-insoluble membrane rafts. We conclude that Rho family GTPases regulate epithelial intercellular junctions via distinct morphological and biochemical mechanisms and that perturbations in barrier function reflect any imbalance in active/resting GTPase levels rather than simply loss or gain of GTPase activity. PMID- 15044154 TI - Calcineurin is required for translational control of protein synthesis in rat pancreatic acini. AB - CCK increases the rate of net protein synthesis in rat pancreatic acini by activating initiation and elongation factors required for translation. The immunosuppressant FK506 inhibits the Ca(2+)-calmodulin-dependent phosphatase calcineurin in pancreatic acinar cells and blocks pancreatic growth induced by chronic CCK treatment. To test a requirement for calcineurin in the activation of the translational machinery stimulated by CCK, we evaluated the effects of FK506 on protein synthesis and on regulatory initiation and elongation factors in rat pancreatic acini in vitro. CCK acutely increased protein synthesis in acini from normal rats with a maximum increase at 100 pM CCK to 170 +/- 11% of control. The immunosuppressant FK506 dose-dependently inhibited CCK-stimulated protein synthesis over the same concentration range that blocked calcineurin activity, as assessed by dephosphorylation of the calcineurin substrate calcium-regulated heat stable protein of 24 kDa. Another immunosuppressant, cyclosporin A, inhibited protein synthesis, but its effects appeared more complex. FK506 also inhibited protein synthesis stimulated by bombesin and carbachol. FK506 did not significantly affect the activity of the initiation factor-2B, or the phosphorylation of the initiation factor-2alpha, ribosomal protein protein S6, or the mRNA cap binding protein eukaryotic initiation factor (eIF) 4E. Instead, blockade of calcineurin with FK506 reduced the phosphorylation of the eIF4E binding protein, reduced the formation of the eIF4F complex, and increased the phosphorylation of eukaryotic elongation factor 2. From these results, we conclude that calcineurin activity is required for protein synthesis, and this action may be related to an effect on the formation of the mRNA cap binding complex and the elongation processes. PMID- 15044155 TI - Estrogen increases retrograde labeling of motoneurons: evidence of a nongenomic mechanism. AB - Estrogen has a variety of neurotrophic effects mediated via different signaling cascades, including ERK and phosphatidylinositol 3-kinase (PI3K) pathways. In this study, we investigated effects of estrogen and inhibitors for ERK and PI3K applied directly onto the cut sciatic nerve on retrograde labeling of lumbar motoneurons. A mix of retrograde tracer (Fluorogold) and 17beta-estradiol, in combination with an antagonist for estrogen receptors ICI 182,780, an inhibitor of ERK1/2 pathway (U0126), an inhibitor of PI3K (LY-294002), or a protein synthesis inhibitor (cycloheximide), was applied to the proximal stump of the transected sciatic nerve for 24 h. Coapplication of Fluorogold with 17beta estradiol produced a significant increase in the number of retrograde-labeled lumbar motoneurons, compared with Fluorogold alone. Estrogen potentiation of retrograde labeling was inhibited by application of ICI 182,780, U0126, LY 294002, and cycloheximide. Immunohistochemical analysis of the sciatic nerve, 24 h following crush injury, revealed accumulation of phospho-ERK in regenerating nerve fibers. The data suggest a role for estrogen, ERK, PI3K, and protein synthesis in the uptake and retrograde transport of Fluorogold. We propose that estrogen action in peripheral nerve fibers is mediated via the ERK and PI3K signaling pathways and is reliant on local protein synthesis. PMID- 15044156 TI - Prostaglandin E2 activates outwardly rectifying Cl(-) channels via a cAMP dependent pathway and reduces cell motility in rat osteoclasts. AB - We examined changes in electrical and morphological properties of rat osteoclasts in response to prostaglandin (PG)E(2). PGE(2) (>10 nM) stimulated an outwardly rectifying Cl(-) current in a concentration-dependent manner and caused a long lasting depolarization of cell membrane. This PGE(2)-induced Cl(-) current was reversibly inhibited by 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB), and tamoxifen. The anion permeability sequence of this current was I(-) > Br(-) approximately Cl(-) > gluconate(-). When outwardly rectifying Cl(-) current was induced by hyposmotic extracellular solution, no further stimulatory effect of PGE(2) was seen. Forskolin and dibutyryl adenosine 3',5'-cyclic monophosphate (DBcAMP) mimicked the effect of PGE(2). The PGE(2)-induced Cl(-) current was inhibited by pretreatment with guanosine 5'-O-2-(thiodiphosphate) (GDPbetaS), Rp-adenosine 3',5'-cyclic monophosphorothioate (Rp-cAMPS), N-(2-[p-bromocinnamylamino]ethyl)-5 isoquinolinesulfonamide dihydrochloride (H-89), and protein kinase A inhibitors. Even in the absence of nonosteoclastic cells, PGE(2) (1 microM) reduced cell surface area and suppressed motility of osteoclasts, and these effects were abolished by Rp-cAMPS or H-89. PGE(2) is known to exert its effects through four subtypes of PGE receptors (EP1-EP4). EP2 and EP4 agonists (ONO-AE1-259 and ONO AE1-329, respectively), but not EP1 and EP3 agonists (ONO-DI-004 and ONO-AE-248, respectively), mimicked the electrical and morphological actions of PGE(2) on osteoclasts. Our results show that PGE(2) stimulates rat osteoclast Cl(-) current by activation of a cAMP-dependent pathway through EP2 and, to a lesser degree, EP4 receptors and reduces osteoclast motility. This effect is likely to reduce bone resorption. PMID- 15044157 TI - The future of academic medicine has begun. PMID- 15044158 TI - Women's health education initiatives: why have they stalled? AB - Since the U.S. Congress first requested an assessment of women's health content in medical school curricula ten years ago, surveys indicate at least a two-fold increase in the number of schools with a women's health curriculum and no change in the number that offer a women's health clinical elective or rotation. Despite a marked increase in the number of schools with an office or program responsible for integration of women's health and gender-specific content into curricula, change has been modest. Reasons for this slow progress include uncertainty about the domain of women's health and what should be included in a curriculum, a lack of practical guidelines for implementation, and institutional resistance to change. The dominant factors that will influence future curriculum development are the increasing scientific knowledge base on sex and gender differences and the emerging scientific field of sex-based biology, both of which have potential to benefit the health of women. Evidence-based data on significant sex and gender differences will provide compelling reasons for schools to integrate this information into curricula, and new educational initiatives must further develop educational models to help implement change. As women's health becomes synonymous with the term "sex and gender differences," the challenge to schools is to address equally in their curricula those unique aspects of women's health that were part of the original intent of the congressional mandate. PMID- 15044159 TI - The "mommy tenure track". AB - For years women have been fighting for equity in the academic work place, but this acceptance comes at a price. Balancing work and family obligations have proven to be an awkward task where maximal satisfaction and gratification in both are not easily attained. Junior-level faculty have the arduous charge of being most productive and prolific exactly during their child-raising years. If our society wants to produce and maintain women professionals, then academic institutions have to be more generous and tolerant in the short-term for a long term gain. The author proposes a "mommy tenure track" in which primary caregivers of infants are allowed to temporarily postpone their tenure clock. She also cites innovative initiatives being carried out at one institution to help women, including young mothers, with their academic careers and promotions. PMID- 15044160 TI - Attitudes of female nurses and female residents toward each other: a qualitative study in one U.S. teaching hospital. AB - PURPOSE: To describe the attitudes of female nurses and female resident physicians toward each other in surgery, internal medicine, obstetrics gynecology, and emergency medicine in one Midwest teaching hospital in the United States. METHOD: Using a qualitative methodology, 51 women were interviewed in 2002: 28 nurses and 23 residents. Questions were asked to determine if and how female nurses and female residents believed gender was a factor in their interprofessional relationships, how each described their relationship with the other, the kind of assistance female nurses provide to female residents, the kind of assistance sought by female residents, and the strengths and challenges of the female nurse-female resident relationship. Data were analyzed using NUD*IST software. RESULTS: Consistent with similar studies conducted in Norway and Australia, the results include the following: For female nurses, occupation is secondary to gender, which is to say that gender is the most important link between female nurses and female residents. For female residents, gender is secondary to occupation/occupational status. CONCLUSIONS: With the number of female residents increasing each year in hospitals, this relationship should be further examined so that dysfunctional communication patterns between the two groups can be challenged. PMID- 15044161 TI - Effects of participation in the Executive Leadership in Academic Medicine (ELAM) program on women faculty's perceived leadership capabilities. AB - PURPOSE: This study measured the impact of participation by women academics in the Executive Leadership in Academic Medicine (ELAM) program as part of a robust evaluation agenda. METHOD: The design is a classic pre/post, within-group, self report study. The survey elicits self-perception about leadership in ten constructs: knowledge of leadership, management, and organizational theory; environmental scanning; financial management; communication; networking and coalition building; conflict management; general leadership; assessment of strengths and weaknesses; acceptance of leadership demands; and career advancement sophistication. The post surveys inquire additionally about perceived program usefulness. Data were collected from 79 participants (1997-98, 1998-99, and 2000-01 classes). Response rates were nearly 100% (pre) and 69% to 76% (post). RESULTS: Statistically significant increases (p <.01) in perceived leadership capabilities were identified across all ten leadership constructs. Gains were large in knowledge of leadership and organizational theory, environmental scanning, financial management, and general leadership. Gains in career building knowledge were large to moderate. More modest were gains in communication, networking, and conflict management. There were significant correlations between each leadership construct and perceived usefulness of the program. CONCLUSIONS: Significant improvements were reported on all leadership constructs, even when participants viewed themselves as already skilled. While it cannot be concluded that participation in ELAM directly and solely caused all improvements, it seems unlikely that midcareer women faculty would improve on all ten constructs in 11 months after program completion by natural maturation alone. Future research will investigate whether the changes are due to ELAM or other factors, and assess whether participants show more rapid advancement into leadership than comparable women not participating in ELAM. PMID- 15044162 TI - Women in academic general surgery. AB - PURPOSE: To portray the professional experiences of men and women in academic general surgery with specific attention to factors associated with differing academic productivity and with leaving academia. METHOD: A 131-question survey was mailed to all female (1,076) and a random 2:1 sample of male (2,152) members of the American College of Surgeons in three mailings between September 1998 and March 1999. Detailed questions regarding academic rank, career aspirations, publication rate, grant funding, workload, harassment, income, marriage and parenthood were asked. A five-point Likert scale measured influences on career satisfaction. Responses from strictly academic and tenure-track surgeons were analyzed and interpreted by gender, age, and rank. RESULTS: Overall, 317 surgeons in academic practice (168 men, 149 women) responded, of which 150 were in tenure track positions (86 men, 64 women). Men and women differed in academic rank, tenure status, career aspirations, and income. Women surgeons had published a median of ten articles compared with 25 articles for men (p <.001). Marriage or parenthood did not influence numbers of publications for women. Overall career satisfaction was high, but women reported feeling career advancement opportunities were not equally available to them as to their male colleagues and feeling isolation from surgical peers. Ten percent to 20% of surgeons considered leaving academia, with women assistant professors (29%) contemplating this most commonly. CONCLUSION: Addressing the differences between men and women academic general surgeons is critical in fostering career development and in recruiting competitive candidates of both sexes to general surgery. PMID- 15044163 TI - Career advancement for women faculty in a U.S. school of medicine: perceived needs. AB - PURPOSE: The percentage of women faculty at the professor level has remained at approximately 11%. The medical community could benefit from knowing what is required to attract, retain, and advance women in the academy. METHOD: The Committee on Women in Medicine and Science at Stanford University School of Medicine was charged with improving career success and well-being of women faculty. In 2001-02, a survey instrument including both needs and perceived school climate was sent to 309 women faculty. Responses were analyzed using one way analyses of variance with Tukey follow-up tests. RESULTS: A total of 163 (53%) faculty responded. The highest ranked needs were a flexible work environment without negative consequences for women with young children (mean = 4.37 on a five-point scale); a three-month sabbatical from clinical and administrative duties (mean = 4.15); departmental mentoring for academic career development (mean = 4.13); and school/departmental administrative secretarial support for grant and manuscript preparation (mean = 4.11). Climate data obtained in 2002, compared to data from similar surveys in 1994 and 1995, showed a nonsignificant decrease in mean ratings for sexual harassment, gender discrimination, and gender insensitivity in the intervening years. Mean ratings for positive climate and cohesion increased between 1994 and 1995 but remained stable from 1995 to 2002. Results of the survey were presented to the dean, faculty, and staff of the medical school. CONCLUSION: Women faculty members were able to clearly indicate specific interventions that would improve their career success and sense of well-being. Since administrators are committed to serious consideration of these recommendations, this was a key step in advancing women's careers in academic medicine at Stanford. PMID- 15044164 TI - The impact of perceived gender bias on obstetrics and gynecology skills acquisition by third-year medical students. AB - PURPOSE: To investigate the perceptions of third-year medical students about how their acquisition of skills during their obstetrics and gynecology clerkship may be affected by their gender. METHOD: From January 1999 to December 2001, all third-year students at one school completing their obstetrics and gynecology rotation were given an anonymous questionnaire addressing whether gender had a positive, negative, or neutral effect on their learning experience. Students were also asked to enumerate procedures they had performed (e.g., deliveries and speculum examinations) and to rate their ability to counsel women on several clinical problems. To further investigate the perceptions of gender discrimination, a focus group of 12 fourth-year students was held. RESULTS: A total of 263 questionnaires (95%) were returned. Of the respondents, 78% of the men felt their gender adversely affected their experience, and 67% of women felt gender had a positive affect. All but five of the remaining students were in the neutral group. Those students who reported a positive gender effect performed significantly more speculum examinations (15.5 versus 12.3), labor coaching (8.7 versus 6.2), and independent deliveries (3.4 versus 2.7) than did the negative gender-effect group. The positive gender-effect group felt more confident of counseling skills. The neutral group did not differ from the negative group. The overall numerical differences among groups were small, and all groups, on average, performed adequate numbers of skills to meet clerkship objectives. CONCLUSIONS: There is a strong perception among medical students that gender influences experience on their obstetrics and gynecology clerkship, but the differences are actually small. Possible reasons for such strong feelings are addressed and related to the history of sexism in reproductive health care and to the ethics of patients' preferences. PMID- 15044166 TI - Protecting America's secrets while maintaining academic freedom. AB - The terrorist attacks of September 11, 2001, and the subsequent anthrax mail attacks, have had a profound impact on Americans' personal and professional lives and have sparked an active debate regarding the delicate balance between the need for national security and the pursuit of academic freedom. Although academic freedom can be defined in many ways, there are four primary tenets of freedom in an academic environment: freedom to research, freedom to publish, freedom to teach, and freedom to speak. Each of these tenets has come under attack in the wake of September 11, 2001. In this report the author further defines academic freedom and reflects upon recent events that have had a real or perceived impact on this freedom, including (1) attempts to categorize and restrict some research as "sensitive," (2) implementation of export control laws and select agent regulations, (3) limitations on the publication of research findings, (4) prohibition of certain foreign nationals from collaborating with U.S. researchers and receiving education and training in U.S. colleges and universities, and (5) restraint of faculty free speech. The author offers some suggestions as to how academia might achieve a proper balance between protecting our national security while promoting and maintaining academic freedom. PMID- 15044167 TI - Lessons learned from a peer review of bedside teaching. AB - While evaluating bedside teaching by attending physicians on the Mayo Clinic's general internal medicine hospital services, the author learned that peer review enhances an understanding of teaching for both observers and subjects of peer review. In this article the author offers five insights derived from his and two colleagues' observations of bedside teaching during a six-month period in 2002. These are (1) the value of peer review to observers, (2) the apparently unlimited number of teaching strategies, (3) the prevalence of missed opportunities to provide feedback to learners, (4) the art of asking questions effectively, and (5) the possible relationship between a teacher's maturity and successful bedside teaching. Regarding the art of asking questions, he encountered four common problems (e.g., the underutilization of questions), but also found that accomplished teachers pursue a course of co-discovery by asking questions alongside their learners. Finally, he learned that experienced attending physicians often demonstrate teaching sessions focused on psychosocial aspects of care, the use of simple questions, and a willingness to expose their own inadequacies. PMID- 15044168 TI - The ethics of cultural competence. AB - Cultural competence curricula have proliferated throughout medical education. Awareness of the moral underpinnings of this movement can clarify the purpose of such curricula for educators and trainees and serve as a way to evaluate the relationship between the ethics of cultural competence and normative Western medical ethics. Though rarely stated explicitly, the essential principles of cultural competence are (1) acknowledgement of the importance of culture in people's lives, (2) respect for cultural differences, and (3) minimization of any negative consequences of cultural differences. Culturally competent clinicians promote these principles by learning about culture, embracing pluralism, and proactive accommodation. Generally, culturally competent care will advance patient autonomy and justice. In this sense, cultural competence and Western medical ethics are mutually supportive movements. However, Western bioethics and the personal ethical commitments of many medical trainees will place limits on the extent to which they will endorse pluralism and accommodation. Specifically, if the values of cultural competence are thought to embrace ethical relativity, inexorable conflicts will be created. The author presents his view of the ethics of cultural competence and places the concepts of cultural competence in the context of Western moral theory. Clarity about the ethics of cultural competence can help educators promote and evaluate trainees' integration of their own moral intuitions, Western medical ethics, and the ethics of cultural competence. PMID- 15044169 TI - Personal illness narratives: using reflective writing to teach empathy. AB - Reflective writing is one established method for teaching medical students empathetic interactions with patients. Most such exercises rely on students' reflecting upon clinical experiences. To effectively elicit, interpret, and translate the patient's story, however, a reflective practitioner must also be self-aware, personally and professionally. Race, gender, and other embodied sources of identity of practitioners and patients have been shown to influence the nature of clinical communication. Yet, although medical practice is dedicated to examining, diagnosing, and treating bodies, the relationship of physicians to their own physicality is vexed. Medical training creates a dichotomy whereby patients are identified by their bodies while physicians' bodies are secondary to physicians' minds. As a result, little opportunity is afforded to physicians to deal with personal illness experiences, be they their own or those of loved ones. This article describes a reflective writing exercise conducted in a second-year medical student humanities seminar. The "personal illness narrative" exercise created a medium for students to elicit, interpret, and translate their personal illness experiences while witnessing their colleagues' stories. Qualitative analysis of students' evaluation comments indicated that the exercise, although emotionally challenging, was well received and highly recommended for other students and residents. The reflective writing exercise may be incorporated into medical curricula aimed at increasing trainees' empathy. Affording students and residents an opportunity to describe and share their illness experiences may counteract the traditional distancing of physicians' minds from their bodies and lead to more empathic and self-aware practice. PMID- 15044170 TI - The moths. PMID- 15044172 TI - The impact of testing accommodations on MCAT scores: descriptive results. AB - PURPOSE: Medical College Admission Test (MCAT) examinees with disabilities who receive accommodations receive flagged scores indicating nonstandard administration. This report compares MCAT examinees who received accommodations and their performances with standard examinees. METHOD: Aggregate history records of all 1994-2000 MCAT examinees were identified as flagged (2,401) or standard (297,880), then further sorted by race/ethnicity (broadly identified as underrepresented minority and non-URM, at the time of testing) and gender. Those with flagged scores were also classified by disability (LD = learning disability, ADHD = attention deficit hyperactivity disorder, LD/ADHD = learning disability and attention deficit hyperactivity disorder, and Other = other disability) and type of accommodation. Mean MCAT scores were calculated for all groups. A group of 866 examinees took the MCAT first as a standard administration and subsequently with accommodations. In a separate analysis, their two sets of scores were compared. RESULTS: Less than 1% of examinees (2,401) had accommodations; of these, 55% were LD, 17% ADHD, 5% LD/ADHD, and 23% Other. Extended time was the most frequently provided accommodation. Mean flagged scores slightly exceeded mean standard scores on all MCAT sections. Examinees who retook the MCAT with accommodations after a standard administration increased their scores by six points, quadrupling the average gain Standard-Standard retest cohort from another study. CONCLUSION: The small but statistically significant different higher flagged scores may reflect either appropriate compensation or overly generous accommodations. Extended time had a positive impact on the scores of those who retested with this accommodation. The validity the flagged MCAT in predicting success in medical school is not known, and further investigation is underway. PMID- 15044173 TI - Guidelines for preclerkship bioterrorism curricula. AB - PURPOSE: To develop medical school curriculum guidelines related to bioterrorism to ensure that future medical graduates are armed with the critical knowledge, skills, and attitudes to face this emerging threat. METHOD: An Internet-based Delphi survey was performed in 2002 under the auspices of the Association of Medical School Microbiology and Immunology Chairs involving 64 medical educators in microbiology, immunology, and infectious diseases representing 54 U.S. medical schools. A 12-member bioterrorism expert reference panel participated in the final phase of the survey. RESULTS: Study participants identified the top educational objectives for the following bioterrorism-related curriculum categories: general issues, biodefense, public health, infection control, infectious diseases, and toxins. CONCLUSION: The study focused on preparedness training through the development of curricular guidelines in predominantly preclerkship medical education by identifying basic science and clinical laboratory aspects of putative biologic weapons organisms and toxins, clinical manifestations of bioterrorist attacks, treatment strategies, epidemiology, and prophylaxis. PMID- 15044174 TI - Upregulation of osteopontin expression is involved in the development of nonalcoholic steatohepatitis in a dietary murine model. AB - The pathogenesis of nonalcoholic steatohepatitis (NASH) is poorly defined. Feeding mice a diet deficient in methionine and choline (MCD diet) induces experimental NASH. Osteopontin (OPN) is a Th1 cytokine that plays an important role in several fibroinflammatory diseases. We examined the role of OPN in the development of experimental NASH. A/J mice were fed MCD or control diet for up to 12 wk, and serum alanine aminotransferase (ALT), liver histology, oxidative stress, and the expressions of OPN, TNF-alpha, and collagen I were assessed at various time points. MCD diet-fed mice developed hepatic steatosis starting after 1 wk and inflammation by 2 wk; serum ALT increased from day 3. Hepatic collagen I mRNA expression increased during 1-4 wk, and fibrosis appeared at 8 wk. OPN protein expression was markedly increased on day 1 of MCD diet and persisted up to 8 wk, whereas OPN mRNA expression was increased at week 4. TNF-alpha expression was increased from day 3 to 2 wk, and evidence of oxidative stress did not appear until 8 wk. Increased expression of OPN was predominantly localized in hepatocytes. Hepatocytes in culture also produced OPN, which was stimulated by transforming growth factor-beta and TNF-alpha. Moreover, MCD diet-induced increases in serum ALT levels, hepatic inflammation, and fibrosis were markedly reduced in OPN(-/-) mice when compared with OPN(+/+) mice. In conclusion, our results demonstrate an upregulation of OPN expression early in the development of steatohepatitis and suggest an important role for OPN in signaling the onset of liver injury and fibrosis in experimental NASH. PMID- 15044175 TI - Regulation of intestinal phosphate cotransporter NaPi IIb by ubiquitin ligase Nedd4-2 and by serum- and glucocorticoid-dependent kinase 1. AB - Serum and glucocorticoid-inducible kinase 1 (SGK1) is highly expressed in enterocytes. The significance of the kinase in regulation of intestinal function has, however, remained elusive. In Xenopus laevis oocytes, SGK1 stimulates the epithelial Na(+) channel by phosphorylating the ubiquitin ligase Nedd4-2, which regulates channels by ubiquitination leading to subsequent degradation of the channel protein. Thus the present study has been performed to explore whether SGK1 regulates transport systems expressed in intestinal epithelial cells, specifically type IIb sodium-phosphate (Na(+)-P(i)) cotransporter (NaPi IIb). Immunohistochemistry in human small intestine revealed SGK1 colocalization with Nedd4-2 in villus enterocytes. For functional analysis cRNA encoding NaPi IIb, the SGK isoforms and/or the Nedd4-2 were injected into X. laevis oocytes, and transport activity was quantified as the substrate-induced current (I(P)). Exposure to 3 mM phosphate induces an I(P) in NaPi IIb-expressing oocytes. Coinjection of Nedd4-2, but not the catalytically inactive mutant (C938S)Nedd4-2, significantly downregulates I(P), whereas the coinjection of (S422D)SGK1 markedly stimulates I(P) and even fully reverses the effect of Nedd4-2 on I(P). The effect of (S422D)SGK1 on NaPi IIb is mimicked by wild-type SGK3 but not by wild-type SGK2, constitutively active (T308D,S473D)PKB, or inactive (K127N)SGK1. Moreover, (S422D)SGK1 and SGK3 phosphorylate Nedd4-2. In conclusion, SGK1 stimulates the NaPi IIb, at least in part, by phosphorylating and thereby inhibiting Nedd4-2 binding to its target. Thus the present study reveals a novel signaling pathway in the regulation of intestinal phosphate transport, which may be important for regulation of phosphate balance. PMID- 15044176 TI - Expression of apical membrane L-glutamate transporters in neonatal porcine epithelial cells along the small intestinal crypt-villus axis. AB - Enteral l-glutamate is extensively utilized as an oxidative fuel by the gut mucosa in the neonate. To identify major uptake pathways and to understand uptake regulation, we examined transport kinetics and molecular identities of apical membrane l-glutamate transporters in epithelial cells sequentially isolated along the small intestinal crypt-villus axis from milk protein-fed, 16-day-old pigs. The distended intestinal sac method was used to isolate 12 sequential cell fractions from the tip villus to the bottom crypt. Initial rates and kinetics of l-glutamate uptake were measured with l-[G-(3)H]glutamate by fast filtration in apical membrane vesicles prepared by Mg(2+) precipitation and differential centrifugation, with membrane potential clamped by SCN(-). Initial l-glutamate uptake results suggested the presence of B(o) and X(AG)(-) transport systems, but the X(AG)(-) system was predominant for uptake across the apical membrane. Kinetic data suggested that l-glutamate uptake through the X(AG)(-) system was associated with higher maximal transport activity but lower transporter affinity in crypt than in villus cells. Molecular identity of the X(AG)(-) glutamate transporter, based on immunoblot and RT-PCR analysis, was primarily the defined excitatory amino acid carrier (EAAC)-1. EAAC-1 expression was increased with cell differentiation and regulated at transcription and translation levels from crypt to upper villus cells. In conclusion, efficiency and capacity of luminal l glutamate uptake across the apical membrane are regulated by changing expression of the X(AG)(-) system transporter gene EAAC-1 at transcription and translation levels as well as maximal uptake activity and transporter affinity along the intestinal crypt-villus axis in the neonate. PMID- 15044177 TI - NADPH oxidase-derived oxidant stress is critical for neutrophil cytotoxicity during endotoxemia. AB - Neutrophils can cause liver injury during endotoxemia through generation of reactive oxygen species. However, the enzymatic source of the oxidant stress and the nature of the oxidants generated remain unclear. Therefore, we investigated the involvement of NADPH oxidase in the pathophysiology by using the NADPH oxidase inhibitor diphenyleneiodonium chloride (DPI) in the galactosamine/endotoxin (700 mg/kg Gal:100 microg/kg ET) model of liver injury. In addition, we measured chlorotyrosine as indicator for hypochlorous acid formation by myeloperoxidase. Gal/ET treatment of male C3HeB/FeJ mice resulted in sinusoidal neutrophil accumulation and parenchymal cell apoptosis (14 +/- 3% of cells) at 6 h. At 7 h, 35% of neutrophils had transmigrated. The number of apoptotic cells increased to 25 +/- 2%, and the overall number of dead cells was 48 +/- 3%; many of them showed the characteristic morphology of necrosis. Hepatocytes, which colocalized with extravasated neutrophils, stained positive for chlorotyrosine and 4-hydroxynonenal (4-HNE) protein adducts. In contrast, animals pretreated with DPI (2.5 mg/kg) were protected against liver injury at 7 h (necrosis = 20 +/- 2%). These livers showed little chlorotyrosine or 4-HNE staining, but apoptosis and neutrophil accumulation and extravasation remained unaffected. However, DPI-treated animals showed serious liver injury at 9 h due to sustained apoptosis. The results indicate that NADPH oxidase is responsible for the neutrophil-derived oxidant stress, which includes formation of hypochlorous acid by myeloperoxidase. Thus NADPH oxidase could be a promising therapeutic target to prevent neutrophil-mediated liver injury. However, the long term benefit of this approach needs to be investigated in models relevant for human liver disease. PMID- 15044179 TI - Activation of cAMP-guanine exchange factor confers PKA-independent protection from hepatocyte apoptosis. AB - cAMP has previously been shown to promote cell survival in a variety of cell types, but the downstream signaling pathway(s) of this antiapoptotic effect is unclear. Thus the role of cAMP signaling through PKA and cAMP-regulated guanine nucleotide exchange factors (cAMP-GEFs) in cAMP's antiapoptotic action was investigated in the present study. cAMP's protective effect against bile acid-, Fas ligand-, and TNF-alpha-induced apoptosis in rat hepatocytes was largely unaffected by the selective PKA inhibitor, Rp-8-(4-chlorophenylthio)-cAMP (Rp cAMP). In contrast, a novel cAMP analog, 8-(4-chlorophenylthio)-2'-O-methyl (CPT 2-Me)-cAMP, which activated cAMP-GEFs in hepatocytes without activating PKA, protected hepatocytes against apoptosis induced by bile acids, Fas ligand, and TNF-alpha. The role of cAMP-GEF and PKA on activation of Akt, a kinase implicated in cAMP survival signaling, was investigated. Inhibition of PKA with RP-cAMP had no effect on cAMP-mediated Akt phosphorylation, whereas CPT-2-Me-cAMP, which did not activate PKA, induced phosphatidylinositol 3-kinase (PI3-kinase)-dependent activation of Akt. Pretreatment of hepatocytes with the PI3-kinase inhibitor, Ly 294002, prevented CPT-2-Me-cAMP's protective effect against bile acid and Fas ligand, but not TNF-alpha-mediated apoptosis. Glucagon, CPT-cAMP, and CPT-2-Me cAMP all activated Rap 1, a downstream effector of cAMP-GEF. These results suggest that a PKA-independent cAMP/cAMP-GEF/Rap pathway exists in hepatocytes and that activation of cAMP-GEFs promotes Akt phosphorylation and hepatocyte survival. Thus a cAMP/cAMP-GEF/Rap/PI3-kinase/Akt signaling pathway may confer protection against bile acid- and Fas-induced apoptosis in hepatocytes. PMID- 15044178 TI - Bax interacts with the voltage-dependent anion channel and mediates ethanol induced apoptosis in rat hepatocytes. AB - Acute ethanol exposure induces oxidative stress and apoptosis in primary rat hepatocytes. Previous data indicate that the mitochondrial permeability transition (MPT) is essential for ethanol-induced apoptosis. However, the mechanism by which ethanol induces the MPT remains unclear. In this study, we investigated the role of Bax, a proapoptotic Bcl-2 family protein, in acute ethanol-induced hepatocyte apoptosis. We found that Bax translocates from the cytosol to mitochondria before mitochondrial cytochrome c release. Bax translocation was oxidative stress dependent. Mitochondrial Bax formed a protein complex with the mitochondrial voltage-dependent anion channel (VDAC). Prevention of Bax-VDAC interactions by a microinjection of anti-VDAC antibody effectively prevented hepatocyte apoptosis by ethanol. In conclusion, these data suggest that Bax translocation from the cytosol to mitochondria leads to the subsequent formation of a Bax-VDAC complex that plays a crucial role in acute ethanol induced hepatocyte apoptosis. PMID- 15044180 TI - Metabolic adjustments with the development, treatment, and recurrence of obesity in obesity-prone rats. AB - Obesity is reaching epidemic proportions and predisposes afflicted individuals to several comorbidities. For these individuals, losing weight has proven to be an easier feat than maintaining a reduced weight. In obesity-prone rats, we examined if there is a metabolic propensity to regain weight after a period of significant weight loss. Twenty-four-hour energy expenditure (EE), sleeping metabolic rate (SMR), and nonprotein respiratory quotient (NPRQ) were obtained by indirect calorimetry with urinary nitrogen analysis and normalized to fat mass (FM) and fat-free mass (FFM) acquired by dual-energy X-ray absorptiometry. Obesity-prone rats were examined after free access to a high-fat diet for 16 wk to establish the obese state. They were again examined after 2 wk of calorie restriction, which reduced body weight (14%) and FM (32%). Rats were again examined after a further 8 wk of intake-regulated weight maintenance or ad libitum feeding that led to weight regain. Metabolic data were compared with preobese and age-matched controls. Weight loss suppressed EE and SMR beyond what was expected for the change in metabolic mass. This elevated metabolic efficiency persisted throughout weight maintenance but resolved after 8 wk of regain. Adjusted NPRQ values were elevated in weight-maintained and weight-regaining rats, suggesting a preference for carbohydrate utilization. These data support the concept that weight reduction in obesity is accompanied by metabolic adjustments beyond the drive to consume calories that predispose to weight regain, and some aspects of this adjustment persist with prolonged weight maintenance and during weight regain. PMID- 15044181 TI - Inhibition of nitric oxide synthase enhances superoxide activity in canine kidney. AB - To evaluate the role of a potential interaction between superoxide anion (O(2)( )) and nitric oxide (NO) in regulating kidney function, we examined the renal responses to intra-arterial infusion of a superoxide dismutase mimetic, tempol (0.5 mg.kg(-1).min(-1)), in anesthetized dogs treated with or without NO synthase inhibitor, N(omega)-nitro-l-arginine (NLA; 50 microg.kg(-1).min(-1)). In one group of dogs (n = 10), tempol infusion alone for 30 min before NLA infusion did not cause any significant changes in renal blood flow (RBF; 5.2 +/- 0.4 to 5.0 +/ 0.4 ml.min(-1).g(-1)), glomerular filtration rate (GFR; 0.79 +/- 0.04 to 0.77 +/ 0.04 ml.min(-1).g(-1)), urine flow (V; 13.6 +/- 2.1 to 13.9 +/- 2.5 microl.min( 1).g(-1)), or sodium excretion (U(Na)V; 2.4 +/- 0.3 to 2.2 +/- 0.3 micromol.min( 1).g(-1)). Interestingly, when tempol was infused in another group of dogs (n = 12) pretreated with NLA, it caused increases in V (4.4 +/- 0.4 to 9.7 +/- 1.4 microl.min(-1).g(-1)) and in U(Na)V (0.7 +/- 0.1 to 1.3 +/- 0.2 micromol.min( 1).g(-1)) without affecting RBF or GFR. Although NO inhibition caused usual qualitative responses in both groups of dogs, the antidiuretic (47 +/- 5 vs. 26 +/- 4%) and antinatriuretic (67 +/- 4 vs. 45 +/- 11%) responses to NLA were seen much less in dogs pretreated with tempol. NLA infusion alone increased urinary excretion of 8-isoprostane (13.9 +/- 2.7 to 22.8 +/- 3.6 pg.min(-1).g(-1); n = 7), which returned to the control levels (11.6 +/- 3.4 pg.min(-1).g(-1)) during coadministration of tempol. These data suggest that NO synthase inhibition causes enhancement of endogenous O(2)(-) levels and support the hypothesis that NO plays a protective role against the actions of O(2)(-) in the kidney. PMID- 15044182 TI - Spinal effects of oxytocin on uterine motility in anesthetized rats. AB - The rat uterus receives an innervation from the lumbosacral and thoracolumbar segments of the spinal cord. These segments receive descending oxytocinergic projections from the paraventricular nucleus of the hypothalamus. We tested the hypothesis that oxytocin regulates uterine motility through a spinal site of action. Oxytocin was administered in anesthetized female rats either intrathecally at the lumbosacral or thoracolumbar spinal cord levels or intravenously. Uterine activity was revealed by measuring changes of intrauterine pressure using an indwelling balloon placed in one caudal uterine horn. The uterus displayed a spontaneous activity characterized by intrauterine pressure rises, the frequency, amplitude, and duration of which were dependent on the stage of the estrous cycle. Oxytocin delivered at the lumbosacral level affected the frequency (during proestrus, estrus, and diestrus) and amplitude (during proestrus and estrus) of uterine activity. During estrus, oxytocin delivered at the thoracolumbar level affected the frequency, amplitude, and duration of the intrauterine pressure rises. Intravenous oxytocin not only affected intrauterine pressure rises (namely amplitude during proestrus and estrus and frequency and duration during estrus) but also increased the basal tone during estrus. The effects of lumbosacral oxytocin were partly mimicked by the oxytocin agonist [Thr(4),Gly(7)]-oxytocin blocked by the oxytocin receptor antagonist atosiban and by hexamethonium. Arginine vasopressin delivered at the lumbosacral level had no effect. These results support our hypothesis that oxytocin released by descending paraventriculo-spinal pathways and acting on spinal oxytocin receptors modulates the activity of the uterus. This regulation is cycle dependent. PMID- 15044183 TI - Two-breath CO(2) test detects altered dynamic cerebrovascular autoregulation and CO(2) responsiveness with changes in arterial P(CO(2)). AB - The new two-breath CO(2) method was employed to test the hypotheses that small alterations in arterial P(CO(2)) had an impact on the magnitude and dynamic response time of the CO(2) effect on cerebrovascular resistance (CVRi) and the dynamic autoregulatory response to fluctuations in arterial pressure. During a 10 min protocol, eight subjects inspired two breaths from a bag with elevated P(CO(2)), four different times, while end-tidal P(CO(2)) was maintained at three levels: hypocapnia (LoCO(2), 8 mmHg below resting values), normocapnia, and hypercapnia (HiCO(2), 8 mmHg above resting values). Continuous measurements were made of mean blood pressure corrected to the level of the middle cerebral artery (BP(MCA)), P(CO(2)) (estimated from expired CO(2)), and mean flow velocity (MFV, of the middle cerebral artery by Doppler ultrasound), with CVRi = BP(MCA)/MFV. Data were processed by a system identification technique (autoregressive moving average analysis) with gain and dynamic response time of adaptation estimated from the theoretical step responses. Consistent with our hypotheses, the magnitude of the P(CO(2))-CVRi response was reduced from LoCO(2) to HiCO(2) [from -0.04 (SD 0.02) to -0.01 (SD 0.01) (mmHg x cm(-1) x s) x mmHg Pco(2)(-1)] and the time to reach 95% of the step plateau increased from 12.0 +/- 4.9 to 20.5 +/- 10.6 s. Dynamic autoregulation was impaired with elevated P(CO(2)), as indicated by a reduction in gain from LoCO(2) to HiCO(2) [from 0.021 +/- 0.012 to 0.007 +/- 0.004 (mmHg x cm(-1) x s) x mmHg BP(MCA)(-1)], and time to reach 95% increased from 3.7 +/- 2.8 to 20.0 +/- 9.6 s. The two-breath technique detected dependence of the cerebrovascular CO(2) response on P(CO(2)) and changes in dynamic autoregulation with only small deviations in estimated arterial P(CO(2)). PMID- 15044184 TI - Evidence that paraventricular nucleus oxytocin neurons link hypothalamic leptin action to caudal brain stem nuclei controlling meal size. AB - Hindbrain projections of oxytocin neurons in the parvocellular paraventricular nucleus (pPVN) are hypothesized to transmit leptin signaling from the hypothalamus to the nucleus of the solitary tract (NTS), where satiety signals from the gastrointestinal tract are received. Using immunocytochemistry, we found that an anorectic dose of leptin administered into the third ventricle (3V) increased twofold the number of pPVN oxytocin neurons that expressed Fos. Injections of fluorescent cholera toxin B into the NTS labeled a subset of pPVN oxytocin neurons that expressed Fos in response to 3V leptin. Moreover, 3V administration of an oxytocin receptor antagonist, [d-(CH2)5,Tyr(Me)2,Orn8] vasotocin (OVT), attenuated the effect of leptin on food intake over a 0.5- to 4 h period (P < 0.05). Furthermore, to determine whether oxytocin contributes to leptin's potentiation of Fos activation within NTS neurons in response to CCK, we counted the number of Fos-positive neurons in the medial NTS (mNTS) after 3V administration of OVT before 3V leptin and intraperitoneal CCK-8 administration. OVT resulted in a significant 37% decrease (P < 0.05) in the potentiating effect of leptin on CCK activation of mNTS neuronal Fos expression. Furthermore, 4V OVT stimulated 2-h food intake by 43% (P < 0.01), whereas 3V OVT at the same dose was ineffective. These findings suggest that release of oxytocin from a descending pPVN-to-NTS pathway contributes to leptin's attenuation of food intake by a mechanism that involves the activation of pPVN oxytocin neurons by leptin, resulting in increased sensitivity of NTS neurons to satiety signals. PMID- 15044185 TI - Expression of NPY Y1 and Y5 receptors in the hypothalamic paraventricular nucleus of aged Fischer 344 rats. AB - Many mammals, nearing the end of life, spontaneously decrease their food intake and body weight, a stage we refer to as senescence. The spontaneous decrease in food intake and body weight is associated with attenuated responses to intracerebroventricular injections of neuropeptide Y (NPY) compared with old presenescent or with young adult rats. In the present study, we tested the hypothesis that this blunted responsiveness involves the number and expression of hypothalamic paraventricular nucleus (PVN) Y(1) and/or Y(5) NPY receptors, both of which are thought to mediate NPY-induced food intake. We found no significant difference in mRNA levels, via quantitative PCR, for Y(1) and Y(5) receptors in the PVN of senescent vs. presenescent rats. In contrast, immunohistochemistry indicated that the number of PVN neurons staining for Y(1) receptor protein was greater in presenescent compared with senescent rats. We conclude that a decreased expression and number of Y(1) or Y(5) receptors in the PVN cannot explain the attenuated responsiveness of the senescent rats to exogenous NPY. PMID- 15044186 TI - Rosuvastatin treatment reverses impaired coronary artery vasodilation in fructose fed, insulin-resistant rats. AB - Insulin resistance (IR) impairs vascular responses in coronary arteries, but mechanisms of dysfunction and approaches to treatment remain unclear. We examined the ability of a new 3-hydroxy-methylglutaryl coenzyme A reductase inhibitor, rosuvastatin, to reverse reduced dilator responses in rats made IR by feeding a fructose-rich diet (FF). Sprague-Dawley rats were randomized to control (normal rat diet) or FF. After 1 wk, rats received rosuvastatin (2 mg/kg) or placebo (saline) subcutaneously for 5 wk. Biochemical measurements and in vitro functional studies of small coronary arteries were performed. Fasting insulin and triglyceride (TG) levels were markedly increased in FF-placebo rats compared with other groups. Rosuvastatin treatment of FF rats normalized TG and modestly decreased insulin levels. ACh-induced dilator responses were depressed in arteries from FF-placebo rats. This impairment was due to decreased responses via calcium-dependent K channels (K(Ca)). Rosuvastatin treatment of FF rats completely reversed the response to ACh to normal levels. Moreover, this recovery in function was due to an improvement in vasodilation via K(Ca). Thus rosuvastatin treatment of IR rats normalizes coronary vascular dilator responses by improving the K(Ca) function. PMID- 15044187 TI - A comparison of adrenergic stress responses in three tropical teleosts exposed to acute hypoxia. AB - Experiments were performed to assess the afferent and efferent limbs of the hypoxia-mediated humoral adrenergic stress response in selected hypoxia-tolerant tropical fishes that routinely experience environmental O(2) depletion. Plasma catecholamine (Cat) levels and blood respiratory status were measured during acute aquatic hypoxia [water Po(2) (Pw(O(2))) = 10-60 mmHg] in three teleost species, the obligate water breathers Hoplias malabaricus (traira) and Piaractus mesopotamicus (pacu) and the facultative air breather Hoplerythrinus unitaeniatus (jeju). Traira displayed a significant increase in plasma Cat levels (from 1.3 +/ 0.4 to 23.3 +/- 15.1 nmol/l) at Pw(O(2)) levels below 20 mmHg, whereas circulating Cat levels were unaltered in pacu at all levels of hypoxia. In jeju denied access to air, plasma Cat levels were increased markedly to a maximum mean value of 53.6 +/- 19.1 nmol/l as Pw(O(2)) was lowered below 40 mmHg. In traira and jeju, Cat release into the circulation occurred at abrupt thresholds corresponding to arterial Po(2) (Pa(O(2))) values of approximately 8.5-12.5 mmHg. A comparison of in vivo blood O(2) equilibration curves revealed low and similar P(50) values (i.e., Pa(O(2)) at 50% Hb-O(2) saturation) among the three species (7.7-11.3 mmHg). Thus Cat release in traira and jeju occurred as blood O(2) concentration was reduced to approximately 50-60% of the normoxic value. Intravascular injections of nicotine (600 nmol/kg) elicited pronounced increases in plasma Cat levels in traira and jeju but not in pacu. Thus the lack of Cat release during hypoxia in pacu may reflect an inoperative or absent humoral adrenergic stress response in this species. When allowed access to air, jeju did not release Cats into the circulation at any level of aquatic hypoxia. The likeliest explanation for the absence of Cat release in these fish was that air breathing, initiated by aquatic hypoxia, prevented Pa(O(2)) values from falling to the critical threshold required for Cat secretion. The ventilatory responses to hypoxia in each species were similar, consisting generally of increases in both frequency and amplitude. These responses were not synchronized with or influenced by plasma Cat levels. Thus the acute humoral adrenergic stress response does not appear to stimulate ventilation during acute hypoxia in these tropical species. PMID- 15044188 TI - Y1- and alpha1-receptor control of basal hindlimb vascular tone. AB - The role of endogenous Y(1)-receptor activation on skeletal muscle vasculature under baseline conditions is currently debated and no in vivo studies have been performed to address this issue. Therefore, this study was designed to address the effect of Y(1)-receptor and/or alpha(1)-adrenoceptor antagonism on basal hindlimb vascular conductance in male Sprague-Dawley rats in vivo. Left hindlimb vascular conductance, carotid artery mean arterial pressure, and heart rate were measured during low volume infusion of N(2)-(diphenylacetyl)-N-[(4 hydroxyphenyl)methyl]-d-arginine amide (BIBP3226; 100 microg/kg), prazosin (20 microg/kg), and combined blockade to the left hindlimb. Vascular conductance increased 1.5 +/- 0.5 microl.min(-1).mmHg(-1) with BIBP3226 infusion, 1.7 +/- 0.5 microl.min(-1).mmHg(-1) with prazosin infusion, and 4.8 +/- 1.0 microl.min( 1).mmHg(-1) with combined blockade (P < 0.05). Interestingly, systolic vascular conductance increased in all three conditions, but diastolic vascular conductance only increased in the two conditions where BIBP3226 was present. These data indicate that Y(1)-receptor activation plays an important role in the regulation of vascular conductance in the resting rat hindlimb. Furthermore, this effect was of the same magnitude as the alpha(1)-adrenoceptor contribution. The differential flow profiles following alpha(1) blockade with and without Y(1)-receptor blockade supports local differences in receptor distribution. PMID- 15044189 TI - Pore loop-mutated rat KIR6.1 and KIR6.2 suppress KATP current in rat cardiomyocytes. AB - Cardiomyocytes express mRNA for all major subunits of ATP-sensitive potassium (K(ATP)) channels: KIR6.1, KIR6.2, SUR1A, SUR2A, and SUR2B. It has remained controversial as to whether KIR6.1 may associate with KIR6.2 to form the tetrameric pore of K(ATP) channels in cardiomyocytes. To explore this possibility, cultured rat cardiomyocytes were examined for an inhibition of K(ATP) current by overexpression of pore loop-mutated (inactive) KIR6.x. Bicistronic plasmids were constructed encoding loop-mutated (AFA or SFG for GFG) rat KIR6.x followed by EGFP. In ventricular myocytes, the overexpression of KIR6.1SFG-pIRES(2)-EGFP or KIR6.2AFA-pIRES(2)-EGFP DNA caused, after 72 h, a major decrease of K(ATP) current density of 85.8% and 82.7%, respectively (P < 0.01), relative to EGFP controls (59 +/- 9 pA/pF). In atrial myocytes, overexpression of these pore-mutated KIR6.x by 6.0-fold and 10.6-fold, as assessed by quantitative immunohistochemistry, caused a decrease of K(ATP) current density of 73.7% and 58.5%, respectively (P < 0.01). Expression of wild type rat KIR6.2 increased the ventricular and atrial K(ATP) current density by 58.3% and 42.9%, respectively (P < 0.01), relative to corresponding EGFP controls, indicating a reserve of SUR to accommodate increased KIR6.x trafficking to the sarcolemma. The results favor the view that KIR6.1 may associate with KIR6.2 to form heterotetrameric pores of native K(ATP) channels in cardiomyocytes. PMID- 15044190 TI - Microvascular dysfunction after transient high glucose is caused by superoxide dependent reduction in the bioavailability of NO and BH(4). AB - We hypothesized that transient high-glucose concentration interferes with mediation by nitric oxide (NO) of flow-induced dilation (FID) of arterioles due to enhanced production of superoxide. In isolated, pressurized (80 mmHg) rat gracilis muscle arterioles ( approximately 130 microm) after transient high glucose treatment (tHG; incubation with 30 mM glucose for 1 h), FID was reduced (maximum: control, 38 +/- 4%; after tHG, 17 +/- 3%), which was not further diminished by the NO synthase (NOS) inhibitor N(omega)-nitro-l-arginine methyl ester (l-NAME; 18 +/- 2%). Correspondingly, an enhanced polyethylene-glycol-SOD (PEG-SOD)-sensitive superoxide production was detected after tHG in carotid arteries by dihydroethydine (DHE) staining. Presence of PEG-SOD during tHG prevented the reduction of FID (41 +/- 3%), which could be inhibited by l-NAME (20 +/- 4%). Administration of PEG-SOD after tHG did not prevent the reduction of FID (22 +/- 3%). Sepiapterin, a precursor of the NO synthase cofactor tetrahydrobiopterin (BH(4)), administered during tHG did not prevent the reduction of FID (maximum, 15 +/- 5%); however, it restored FID when administered after tHG (32 +/- 4%). Furthermore, inhibition of either glycolysis by 2 deoxyglucose or mitochondrial complex II by 2-thenoyltrifluoroacetone reduced the tHG-induced DHE-detectable enhanced superoxide production in carotid arteries and prevented FID reduction in arterioles (39 +/- 5 and 35 +/- 2%). Collectively, these findings suggest that in skeletal muscle arterioles, a transient elevation of glucose via its increased metabolism, elicits enhanced production of superoxide, which decreases the bioavailability of NO and the level of the NOS cofactor BH(4), resulting in a reduction of FID mediated by NO. PMID- 15044191 TI - Cardioprotection by glucose-insulin-potassium: dependence on KATP channel opening and blood glucose concentration before ischemia. AB - We tested the hypothesis that glucose-insulin-potassium (GIK)-induced protection against myocardial infarction depends on ATP-dependent K(+) (K(ATP)) channel activation and is abolished by hyperglycemia before the ischemia. Dogs were subjected to a 60-min coronary artery occlusion and 3-h reperfusion in the absence or presence of GIK (25% dextrose; 50 IU insulin/l; 80 mM/l KCl infused at 1.5 ml x kg(-1) x h(-1)) beginning 75 min before coronary artery occlusion or 5 min before reperfusion. The role of K(ATP) channels was evaluated by pretreatment with glyburide (0.1 mg/kg). The efficacy of GIK was investigated with increases in blood glucose (BG) concentrations to 300 or 600 mg/dl or experimental diabetes (alloxan/streptozotocin). Infarct size (IS) was 29 +/- 2% of the area at risk in control experiments. GIK decreased (P < 0.05) IS when administered beginning 5 min before reperfusion. This protective action was independent of BG (13 +/- 2 and 12 +/- 2% of area at risk; BG = 80 or 600 mg/dl, respectively) but was abolished in dogs receiving glyburide (30 +/- 4%), hyperglycemia before ischemia (27 +/- 4%), or diabetes (25 +/- 3%). IS was unchanged by GIK when administered before ischemia independent of BG (31 +/- 3, 27 +/- 2, and 35 +/- 3%; BG = 80, 300, and 600 mg/dl, respectively). The insulin component of GIK promotes cardioprotection by K(ATP) channel activation. However, glucose decreases K(ATP) channel activity, and this effect predominates when hyperglycemia is present before ischemia. PMID- 15044192 TI - Attenuation of neointima formation through the inhibition of DNA repair enzyme PARP-1 in balloon-injured rat carotid artery. AB - Increased oxidative stress is a major characteristic of restenosis after angioplasty. The oxidative stress is mainly created by oxidants such as reactive oxygen species (ROS), which are assumed to play an important role in neointima formation after angioplasty. DNA is a sensitive target for oxidants; however, oxidative DNA damage remains a poorly examined field in the pathogenesis of restenosis. In the present study, we demonstrated that the expression of the oxidative DNA damage marker 7,8-dihydro-8-oxo-2'-deoxyguanosine (8-oxo-dG) was quickly increased in rat carotid arteries after balloon injury. It reached its peak at 14 days after injury and still kept high expression at 28 days after injury. The immunostaining of 8-oxo-dG was present predominantly in the neointima. In response to oxidative DNA damage, the DNA repair enzyme poly(ADP ribose) polymerase-1 (PARP-1) was significantly increased after balloon injury. The time course change and location of PARP-1 is similar to that of 8-oxo-dG. Daily injections of the PARP-1 inhibitor PJ34 (5 mg.kg(-1).day(-1) ip) attenuated neointima formation by approximately 40% at 7, 14, and 28 days after balloon injury. Treatment with PJ34 inhibited leukocyte infiltration and improved both anatomic (reendothelialization) and functional (endothelial function) recovery of endothelial cells after balloon injury. In conclusion, levels of oxidative DNA damage and the DNA repair enzyme PARP-1 are increased in vessels after balloon injury. Inhibition of PARP-1 attenuates neointima formation through inhibition of leukocyte infiltration and improvement of endothelial cell recovery after balloon injury. Targeting of the DNA repair enzyme might be a therapeutic strategy for restenosis. PMID- 15044193 TI - Aging increases stiffness of cardiac myocytes measured by atomic force microscopy nanoindentation. AB - It is well established that the aging heart exhibits left ventricular (LV) diastolic dysfunction and changes in mechanical properties, which are thought to be due to alterations in the extracellular matrix. We tested the hypothesis that the mechanical properties of cardiac myocytes significantly change with aging, which could contribute to the global changes in LV diastolic dysfunction. We used atomic force microscopy (AFM), which determines cellular mechanical property changes at nanoscale resolution in myocytes, from young (4 mo) and old (30 mo) male Fischer 344 x Brown Norway F1 hybrid rats. A measure of stiffness, i.e., apparent elastic modulus, was determined by analyzing the relationship between AFM indentation force and depth with the classical infinitesimal strain theory and by modeling the AFM probe as a blunted conical indenter. This is the first study to demonstrate a significant increase (P < 0.01) in the apparent elastic modulus of single, aging cardiac myocytes (from 35.1 +/- 0.7, n = 53, to 42.5 +/- 1.0 kPa, n = 58), supporting the novel concept that the mechanism mediating LV diastolic dysfunction in aging hearts resides, in part, at the level of the myocyte. PMID- 15044194 TI - Exogenous NO triggers preconditioning via a cGMP- and mitoKATP-dependent mechanism. AB - Exogenous nitric oxide (NO) triggers a preconditioning-like effect in heart via a pathway that is dependent on reactive oxygen species. This study examined the signaling pathway by which the NO donor S-nitroso-N-acetylpenicillamine (SNAP, 2 microM) triggers its anti-infarct effect. Isolated rabbit hearts experienced 30 min of regional ischemia and 120 min of subsequent reperfusion. Infarct size was determined by triphenyltetrazolium chloride staining. Infarct size was reduced from 30.5 +/- 3.0% of the risk zone in control hearts to 10.2 +/- 2.0% in SNAP treated hearts. Bracketing the SNAP infusion with either the guanylyl cyclase blocker 1H-[1,2,4]oxadiazole[4,3-a]quinoxalin-1-one (2 microM) or the mitochondrial ATP-sensitive K(+) (mitoK(ATP)) channel blocker 5-hydroxydecanoate (200 microM) completely blocked the infarct-sparing effect of SNAP (34.3 +/- 3.8 and 32.2 +/- 1.6% infarction, respectively). Pretreatment of hearts with 8-(4 chlorophenylthio)-guanosine 3',5'-cyclic monophosphate (10 microM), which is a cell-permeable cGMP analog that activates protein kinase G, mimicked the preconditioning effect of SNAP by reducing infarct size to 7.5 +/- 1.1% of the risk zone. This salutary effect was abolished by either the free radical scavenger N-(2-mercaptopropionyl)glycine (1 mM) or 5-hydroxydecanoate (100 microM; 28.9 +/- 2.7 and 33.6 +/- 5.0% infarction of the risk zone, respectively). To confirm these functional data and the effect of SNAP on the guanylyl cyclase-protein kinase G signaling pathway, cGMP levels were measured. SNAP increased the level from 0.18 +/- 0.04 to 0.61 +/- 0.14 pmol/mg of protein (P < 0.05). These data suggest that exogenous NO triggers the preconditioning effect by initiating a cascade of events including stimulation of guanylyl cyclase to make cGMP, activation of protein kinase G, opening of mitoK(ATP) channels, and, finally, production of reactive oxygen species. PMID- 15044195 TI - A (13)C isotopomer kinetic analysis of cardiac metabolism: influence of altered cytosolic redox and [Ca(2+)](o). AB - Rat hearts were perfused with mixtures of [3-(13)C]pyruvate and [3-(13)C]lactate (to alter cytosolic redox) at low (0.5 mM) or high (2.5 mM) Ca(2+) concentrations to alter contractility. Hearts were frozen at various times after exposure to these substrates, were extracted, and were then analyzed by (13)C NMR spectroscopy. The time-dependent multiplets observed in the (13)C NMR resonances of glutamate in all hearts and in malate and aspartate in hearts perfused with high-pyruvate/low-lactate concentrations were analyzed using a kinetic model of the tricarboxylic acid (TCA) cycle. The analysis showed that TCA cycle flux (V(TCA)) and exchange flux (V(X)) that involved cycle intermediates were both sensitive to cell redox and altered Ca(2+) concentration, and the ratio of these fluxes (V(X)/V(TCA)) varied >10-fold. PMID- 15044196 TI - Role of adhesion and contraction in Rac 1-regulated endothelial barrier function in vivo and in vitro. AB - We demonstrated previously that inhibition of the small GTPase Rac-1 by Clostridium sordellii lethal toxin (LT) increased the hydraulic conductivity (L(p)) of rat venular microvessels and induced gap formation in cultured myocardial endothelial cells (MyEnd). In MyEnd cells, we also demonstrated that both LT and cytochalasin D reduced cellular adhesion of vascular endothelial (VE) cadherin-coated beads. Here we further evaluate the contribution of actin depolymerization, myosin-based contraction, and VE-cadherin linkage to the actin cytoskeleton to LT-induced permeability. The actin-depolymerizing agent cytochalasin D increased L(p) in single rat mesenteric microvessels to the same extent as LT over 80 min. However, whereas the actin-stabilizing agent jasplakinolide blunted the L(p) increase due to cytochalasin D by 78%, it had no effect on the LT response. This conforms to the hypothesis that the predominant mechanism whereby Rac-1 stabilizes the endothelial barrier in intact microvessels is separate from actin polymerization and likely at the level of the VE-cadherin linkage to the actin cytoskeleton. In intact vessels, neither inhibition of contraction (butanedione monoxime, an inhibitor of myosin ATPase) nor inhibition of Rho kinase (Y-27632) modified the response to LT, even though both inhibitors lowered resting L(p). In contrast butanedione monoxime and inhibition of myosin light chain kinase completely inhibited LT-induced intercellular gap formation and largely reduced the LT-induced permeability increase in MyEnd monolayers. These results support the hypothesis that the contractile mechanisms that contribute to the formation of large gaps between cultured endothelial cells exposed to inflammatory conditions do not significantly contribute to increased permeability in intact microvessels. PMID- 15044197 TI - Chronic AT(1) receptor blockade alters mechanisms mediating responses to hypoxia in rat skeletal muscle resistance arteries. AB - The goal of this study was to determine the effect of angiotensin type 1 (AT(1)) receptor antagonism on vasodilator responses in isolated skeletal muscle resistance arteries. Normotensive Sprague-Dawley rats were fed normal rat chow with the AT(1) receptor antagonist losartan (1mg/ml) in the drinking water for 7 days and compared with untreated control rats. Changes in the diameter of isolated resistance arteries supplying the gracilis muscle were assessed with a video micrometer. Arteriolar responses to acetylcholine, iloprost, and sodium nitroprusside were unaffected by losartan administration, whereas dilation to reduced Po(2) was converted into a constriction. Hypoxia-induced constriction of vessels from losartan-treated rats was inhibited by endothelium removal or indomethacin (1 microM). Blockade of the PGH(2)-thromboxane A(2) receptor with SQ 29548 (10 microM), thromboxane synthase inhibition with dazoxiben (10 microM), or the addition of the superoxide dismutase mimetic 4-hydroxy-2,2,6,6 tetramethylpiperidine 1-oxyl (TEMPOL, 100 microM) converted hypoxic vasoconstriction to a dilation that was blocked by inhibiting nitric oxide synthase with N(omega)-nitro-l-arginine methyl ester (100 microM). These data suggest that AT(1) receptor activation has an important role in maintaining the vascular release of prostaglandins responsible for mediating hypoxic dilation in skeletal muscle microvessels. PMID- 15044198 TI - Bone marrow stromal cells improve cardiac performance in healed infarcted rat hearts. AB - Postinfarct congestive heart failure is one of the leading causes of morbidity and mortality in developed and developing countries. The main purpose of this study was to investigate whether transplantation of bone marrow stromal cells (BMSC) directly into the myocardium could improve the performance of healed infarcted rat hearts. Cell culture medium with or without BMSC was injected into borders of cardiac scar tissue 4 wk after experimental infarction. Cardiac performance was evaluated 2 wk after cellular (n = 10) or medium (n = 10) injection by electro- and echocardiography. Histological study was performed 3 wk after treatment. Electrocardiography of BMSC-treated infarcted rats showed electrical and mechanical parameters more similar to those in control than in medium-treated animals: a normal frontal QRS axis in 6 of 10 BMSC-treated and all control rats and a rightward deviation of the QRS axis in all 10 medium-treated animals. BMSC treatment, assessed by echocardiography, improved fractional shortening (39.00 +/- 4.03%) compared with medium-treated hearts (18.20 +/- 0.74%) and prevented additional changes in cardiac geometry. Immunofluorescence microscopy revealed colocalization of 4',6-diamidino-2-phenylindole-labeled nuclei of transplanted cells with cytoskeletal markers for cardiomyocytes and smooth muscle cells, indicating regeneration of damaged myocardium and angiogenesis. These data provide strong evidence that BMSC implantation can improve cardiac performance in healed infarctions and open new promising therapeutic opportunities for patients with postinfarction heart failure. PMID- 15044199 TI - Delta-opioid receptor agonist reduces severity of postresuscitation myocardial dysfunction. AB - Postresuscitation myocardial dysfunction is recognized as a leading cause of early death after initially successful cardiopulmonary resuscitation (CPR). In the present study, we hypothesized that a delta-opioid receptor agonist would decrease the severity of postresuscitation myocardial dysfunction and improve survival. Fifteen Sprague-Dawley rats, fasted overnight with access to water, were anesthetized by an injection of 45 mg/kg ip pentobarbital sodium. Additional doses of 10 mg/kg were administered at hourly intervals but not within 30 min before induced ventricular fibrillation (VF). Either the delta-opioid receptor agonist pentazocine (300 microg/kg), pentazocine pretreated with the opioid receptor-blocking agent naloxone (1 mg/kg), or saline placebo was injected into the right atrium after 5 min of untreated VF and 3 min before initiation of CPR. After an additional 8 min of CPR administration, defibrillation was attempted. All animals were successfully resuscitated. Left ventricular rate of pressure increase at 40 mmHg and cardiac index values were significantly improved in pentazocine-treated animals, which also had significantly longer survival times (60 +/- 11 vs. 16 +/- 7 h; P < 0.01). Except for ease of defibrillation, the beneficial effects of pentazocine were completely abolished by pretreatment with naloxone. The concept of pharmacological hibernation employing a delta-opioid receptor agonist is a novel and promising intervention for minimizing global ischemic injury during CPR and postresuscitation myocardial dysfunction. PMID- 15044200 TI - Embolization itself stimulates thrombus propagation in pulmonary embolism. AB - The role of active thrombosis in the pathophysiology of pulmonary embolism is unclear. We tested the hypothesis that venous thrombi significantly increase their thrombotic activity once they embolize into the high-flow circulation of the pulmonary arteries. Thrombotic activity was measured using an immunoassay that measures both fibrinopeptide B (FPB) as well as its most abundant metabolite des-arginine FPB. Thrombi were formed in the femoral veins of adult dogs. In one group, the thrombi were embolized without anticoagulation. In the second group, heparin (300 U/kg bolus, then 90 U x kg(-1) x h(-1) infusion) was administered before embolization to prevent subsequent thrombotic activity. Plasma FPB concentrations were significantly suppressed in the heparinized group relative to the nonheparinized group for 1 h postembolization (P = 0.038). We conclude that pulmonary embolization itself causes preexisting venous thrombi to greatly intensify their thrombotic activity and that embolization-associated thrombus propagation can be prevented by heparin. PMID- 15044201 TI - Burkholderia cenocepacia lipopolysaccharide, lipid A, and proinflammatory activity. AB - Organisms from the Burkholderia cepacia complex are important pathogens in cystic fibrosis and are associated with increased rates of sepsis and death. These organisms comprise nine closely related species known as genomovars. B. cenocepacia (genomovar III) is the most prevalent and appears the most virulent. We investigated the biological activity of a reference panel of strains using whole-cell lysates to induce septic-shock related cytokines from differentiated human monocytic cells. We found varying biological activity within and between genomovars, with B. cenocepacia strains possessing the greatest cytokine induction activity. This activity was CD-14 dependent, suggesting that LPS was responsible for the cytokine induction. Cytokine induction was not simply related to the expression of rough or smooth LPS. We purified LPS from two strains, B. cenocepacia LMG 12614 and B. multivorans LMG 14273, each possessing rough LPS. Divergence in biological activity of the two genomovars was preserved when human monocytic cells were stimulated with purified LPS. Lipid A purified from LMG 14273 and LMG 12614 were analyzed by matrix-assisted laser desorption ionization/time of flight mass spectrometry. Lipid A from the less effective cytokine inducer LMG 14273 was found to be missing a beta-hydroxymyristate (3-OH C14:0) relative to the lipid A of B. cenocepacia LMG 12614. PMID- 15044202 TI - Mass spectrometric analysis of surfactant metabolism in human volunteers using deuteriated choline. AB - Surfactant reduces surface tension at pulmonary air-liquid interfaces. Although its major component is dipalmitoyl-phosphatidylcholine (PC16:0/16:0), other PC species, principally palmitoylmyristoyl-PC, palmitoylpalmitoleoyl-PC, and palmitoyloleoyl-PC, are integral components of surfactant. The composition and metabolism of PC species depend on pulmonary development, respiratory rate, and pathologic alterations, which have largely been investigated in animals using radiolabeled precursors. Recent advances in mass spectrometry and availability of precursors carrying stable isotopes make metabolic experiments in human subjects ethically feasible. We introduce a technique to quantify surfactant PC synthesis in vivo using deuteriated choline coupled with electrospray ionization tandem mass spectrometry. Endogenous PC from induced sputa of healthy volunteers comprised 54.0 +/- 1.5% PC16:0/16:0, 9.7 +/- 0.7% palmitoylmyristoyl-PC, 10.0 +/- 1.0% palmitoylpalmitoleoyl-PC, and 13.1 +/- 0.3% palmitoyloleoyl-PC. Infusion of deuteriated choline chloride (3.6 mg/kg body weight) over 3 hours resulted in linear incorporation into PC over 30 hours. After a plateau of 0.61 +/- 0.04% labeled PC between 30 and 48 hours, incorporation decreased to 0.30 +/- 0.02% within 7 days. Compared with native PC, fractional label was initially lower for PC16:0/16:0 (31.9 +/- 8.3%) but was higher for palmitoyloleoyl-PC (21.0 +/- 1.2%), and equilibrium was achieved after only 48 hours. We conclude that infusion of deuteriated choline and electrospray ionization tandem mass spectrometry is useful to investigate surfactant metabolism in humans in vivo. PMID- 15044203 TI - Role of microvascular permeability on physiologic differences in asthma and eosinophilic bronchitis. AB - Asthma and eosinophilic bronchitis are characterized by a similar type of eosinophilic inflammation. However, eosinophilic bronchitis differs from asthma in that there is no variable airflow obstruction or airway hyperresponsiveness. We evaluated the roles of vascular endothelial growth factor (VEGF) and microvascular permeability in causing these differences between the two diseases. Inflammatory indexes in induced sputum, exhaled nitric oxide levels, and vascular permeability index were examined in 11 normal control subjects, 19 beclomethasone dipropionate (BDP)-treated subjects with asthma, 20 non-BDP-treated subjects with asthma, and 17 patients with eosinophilic bronchitis. The percentage of eosinophils in sputum and exhaled nitric oxide levels were significantly higher in non-BDP-treated subjects with asthma and patients with eosinophilic bronchitis than in other two groups; however, VEGF levels and vascular permeability index were significantly higher in non-BDP-treated (VEGF: mean; 4,710 [SD; 1,150] pg/ml, p < 0.0001; vascular permeability index: 0.028 [0.009], p < 0.0001) and BDP-treated (2,560 [1,070] pg/ml, p = 0.0002; 0.016 [0.006], p = 0.004) subjects with asthma than in patients with eosinophilic bronchitis (1,120 [800] pg/ml; 0.01 [0.005]) and normal control subjects (1,390 [1,280] pg/ml; 0.008 [0.003]). We found significant correlations between the VEGF level and the airway vascular permeability index in all patient groups. Thus, interaction between airway microcirculation and VEGF may be a key element in differences in airway function between asthma and eosinophilic bronchitis. PMID- 15044204 TI - Hypoxia causes glucose intolerance in humans. AB - Hypoxic respiratory diseases are frequently accompanied by glucose intolerance. We examined whether hypoxia is a cause of glucose intolerance in healthy subjects. In a double-blind within-subject crossover design, hypoxic versus normoxic conditions were induced in 14 healthy men for 30 minutes by decreasing oxygen saturation to 75% (versus 96% in control subjects) under the conditions of a euglycemic clamp. The rate of dextrose infusion needed to maintain stable blood glucose levels was monitored. Neurohormonal stress response was evaluated by measuring catecholamine and cortisol concentrations as well as cardiovascular parameters, and symptoms of anxiety. To differentiate between the effects of stress hormonal response, and hypoxia itself, on glucose intolerance, we performed hypoglycemic clamps as a nonspecific control. We found a significant decrease in dextrose infusion rate over a period of 150 minutes after the start of hypoxia (p < 0.01). Hypoxia also increased plasma epinephrine concentration (p < 0.01), heart rate (p < 0.01), and symptoms of anxiety (p < 0.05), whereas the other parameters remained unaffected. Glucose intolerance was closely comparable between hypoxic and hypoglycemic conditions (p < 0.9) despite clear differences in stress hormonal responses. Hypoxia acutely causes glucose intolerance. One of the factors mediating this effect could be an elevated release of epinephrine. PMID- 15044205 TI - Effects of interferon-gamma 1b on biomarker expression in patients with idiopathic pulmonary fibrosis. AB - In a recent study of IFN-gamma 1b in 330 patients with idiopathic pulmonary fibrosis (IPF), progression-free survival was unchanged; however, a trend toward lower mortality was seen in IFN-gamma 1b-treated patients compared with placebo treated patients (9.9 vs. 16.7%; p = 0.08). The purpose of this randomized, double-blind, placebo-controlled trial was to characterize molecular effects of subcutaneous IFN-gamma 1b (200 microg) thrice weekly for 6 months versus placebo in 32 patients with IPF. Messenger RNA in transbronchial lung biopsies and bronchoalveolar lavage cell pellet and protein levels in bronchoalveolar lavage fluid (BALF) and plasma were evaluated. After IFN-gamma 1b treatment, IFN inducible T cell-alpha chemoattractant/CXCL11 (a chemokine with immunomodulatory, antiangiogenic, and defensin-like antimicrobial properties) increased in BALF (p = 0.016) and plasma (p < 0.001); BALF levels of epithelial neutrophil-activating protein-78/CXCL5 (p = 0.054), platelet-derived growth factor A (p = 0.033), and Type I procollagen (p = 0.096) were lower; and IFN-gamma levels were higher (p = 0.093) versus placebo. For messenger RNA in transbronchial biopsies, trends (p > 0.05 and